Detergent composition

ABSTRACT

The present invention relates to a detergent composition comprising an endo-glucanase that provides I improved detergency performance. The invention also relates to a detergent composition comprising a combination of an endo-glucanase and other enzymes. One aspect of the invention relates to a process of washing a fabric of hard surface with the endo-glucanase detergent.

The present invention relates to a detergent composition comprising anendo-glucanase that provides improved detergency performance. Theinvention also relates to a detergent composition comprising acombination of an endo-glucanase and other enzymes. One aspect of theinvention relates to a process of washing a fabric or hard surface withthe endo-glucanase containing detergent.

BACKGROUND OF THE INVENTION

An important objective of a washing process is to transfer, ascompletely as possible, the soil from the object being washed into thewashing solution, such that the soil can then be discarded with the washsolution. A wash process which merely redistributes the soil on theobject being washed is obviously not satisfactory. Thus evaluations ofdetergency performance, i.e. the performance of washing processes, mustinclude consideration both of the removal of soil from the object beingwashed and of the redeposition of this soil onto the object being washedor onto the equipment being used for the wash process.

With some soils and some surfaces it is very difficult to achieve asatisfactory detergency performance. Problem soils include particulatesoils such as clays and carbon. Visible soils are often difficult towash away completely because they are bound to, or attracted to, asurface by traces of non-visible, sticky substances. Problem surfacesinclude the surfaces of cotton fibres.

To help overcome such problem soils, detergent formulations may includean anti-redeposition agent. The anti-redeposition agent is added inorder to ensure that the above mentioned problem soils, once detachedfrom the fabrics, can be kept dissolved or suspended in the wash liquorin such a way that they are not re-deposited on the cleaned fabric.

To obtain both an anti-redeposition effect and a cleaning effect it hasbeen suggested to add a mixture of cellulases to the detergent, onecellulase having anti-redeposition effect and one having acellulose-degrading effect. EP 822 973 relates to a detergentcomposition comprising a mixture of cellulases and optionally alsocontaining additional enzymes. However, there is a need for improvedcombinations of enzymes having anti-redeposition effect and otherenzymes, such as proteases, amylases, hemi-cellulases, lipases andpectinases.

Enzymes are commonly used to remove the sticky substances that increasesoil binding. There is a need for improved enzyme performance in termsof soil removal and/or prevention of soil redeposition.

SUMMARY OF THE INVENTION

The invention relates to a detergent composition comprising ananti-redeposition endo-glucanase, i.e. an endo-glucanase havinganti-redeposition effect. The inventors have found that suchendo-glucanase containing detergents give advantages when used forwashing fabrics (especially laundry washing) or hard surfaces(especially automatic dish washing).

In the context of the present invention, an endo-glucanase havinganti-redeposition effect is characterised by its ability to preventredeposition in a wash test.

The inventors also have found that detergent compositions comprisingspecific combinations of certain endo-glucanase(s) havinganti-redeposition effect and certain cellulase(s) having increasedstability towards anionic tensides such as linear (straight-chain) alkylbenzene sulfonates (often referred to as “LAS”), have advantagescompared to the prior art detergent compositions described in EP 822973. For instance, a decreased amount of enzyme protein is necessary toobtain the desired cleaning and anti-redeposition effect. This resultsin improved product economy.

The inventors also have found that it is surprisingly advantageous toinclude an endo-glucanase having anti-redeposition effect in detergentsthat also contain other enzymes. These other enzymes include, forexample, enzymes that are classified as protease, amylase,beta-glucanase, lipase, hemi-cellulase, cutinase, pectinase and pectatelyase.

Thus in a first aspect the invention relates to a detergent compositioncomprising an endo-glucanase, wherein the endo-glucanase is selectedfrom one of:

-   (i) the endo-glucanase having the amino acid sequence of position 1    to position 773 of SEQ ID NO: 2;-   (ii) an endo-glucanase having a sequence of at least 90% identity to    the amino acid sequence of position 1 to position 773 of SEQ ID    NO:2; or a fragment thereof that has endo-glucanase activity;-   (iii) an endo-glucanase characterized by the wash test method    disclosed herein.

In a second aspect the invention relates to a detergent compositioncomprising anionic tensides and a combination of an endo-glucanase asdefined above and a fungal cellulase, wherein both enzymes are stable inthe presence of anionic tensides.

In a third aspect the invention relates to a detergent compositioncontaining both an endo-glucanase as defined above and one or more otherenzyme from classes such as, but not limited to protease, amylase,beta-glucanase, lipase, hemi-cellulase, cutinase, pectinase and pectatelyase.

In an aspect the invention relates to a process of using a detergentcomposition of the invention for washing or cleaning of fabric, hardsurfaces or other objects in need of cleaning.

DETAILED DESCRIPTION OF THE INVENTION

The invention relates to a detergent composition comprising anendo-glucanase having anti-redeposition effect.

The Endo-Glucanase with Anti-Redeposition Effect

The endo-glucanase having anti-redeposition effect is according to theinvention selected from one of:

-   (i) the endo-glucanase having the amino acid sequence of position 1    to position 773 of SEQ ID NO: 2;-   (ii) an endo-glucanase having a sequence of at least 90% identity to    the amino acid sequence of position 1 to position 773 of SEQ ID    NO:2; or a fragment thereof that has endo-glucanase activity;-   (iii) any endo-glucanase characterized by the wash test method    disclosed below.

In a preferred embodiment the endo-glucanase is derived from Bacillussp. M349, DSM 12648 and also shown in SEQ ID NO: 2 herein or a sequencebeing 90% identical thereto.

The strain Bacillus sp. M349, which has been isolated from a soil sampleoriginating in Greece, was deposited by the inventors according to theBudapest Treaty on the International Recognition of the Deposit ofMicroorganisms for the Purposes of Patent Procedure at the DeutscheSammiung von Mikroorganismen und Zellkulturen GmbH, Mascheroder Weg lb,D-38124 Braunschweig, Germany, on 25 Jan. 1999 under the depositionnumber DSM 12648.

The deposit was made by Novo Nordisk A/S and was later assigned toNovozymes A/S.

Detergent Compositions Comprising Anionic Tensides

A preferred embodiment the invention relates to a detergent compositioncomprising anionic tensides, and to a combination of an endo-glucanaseas defined above and a fungal cellulase, wherein both enzymes are stablein the presence of anionic tensides, such as LAS.

For the purpose of the present invention, enzymes that are stable in LASare defined by a LAS stability test. Enzymes that give a LAS stabilityresult of at least 50% are regarded as being LAS stable.

Detergents commonly contain more than one type of surfactant, forexample combiations of both anionic and nonionic surfactants. In apreferred embodiment the invention reates to detergent formulations inwhich the ratio (based on weight) between anionic and nononicsurfactants is >1:1, preferably >1.5:1.

Thus, in a preferred embodiment the invention relates to a detergentcomposition herein

-   (a) the endo-glucanase is selected from one of:    -   (i) the endo-glucanase having the amino acid sequence of        position 1 to position 773 of SEQ ID NO: 2;    -   (ii) an endo-glucanase having a sequence of at least 90%        identity to the amino acid sequence of position 1 to position        773 of SEQ ID NO:2; or a fragment thereof that has        endo-glucanase activity; and-   (b) the cellulase is selected from one of:    -   (i) the cellulase having the amino acid sequence of position 1        to position 299 of SEQ ID NO: 4; or    -   (ii) a cellulase having a sequence of at least 70% identity to        the amino acid sequence of position 1 to position 299 of SEQ ID        NO:4, or a fragment thereof that has cellulase activity.

Alternatively, the endo-glucanase of (a)(i) above is derived fromBacillus sp. KSMS237 deposited as FERM P-16067 (with the PatentMicroorganism Depository, National Institute of Bioscience andHuman-Technology, Agency of Industrial Science & Industry, Tsukuba-shi,Ibaraki, 305 Japan) and shown in position 1 to 824 of SEQ ID NO: 1 of JP2000210081 A (hereby incorporated by reference).

Other alternative cellulases are the LAS resistant variants disclosed inExample 8 of WO 98/12307. Preferred embodiments of the inventioncomprise these alternative cellulases in combination with theendo-glucanases of (a) above.

In a preferred embodiment the cellulase is a Thielavia terrestriscellulase, preferably the cellulase disclosed in SEQ ID NO: 9 in WO96/29397 and SEQ ID NO: 4 herein or an enzyme with at least 70% identitythereto. In a preferred embodiment the cellulase is the Thielaviaterrestris variant disclosed in Example 1 of WO 98/12307.

Identity

In the context of the present invention the degree of identity isdetermined between two sequences indicating a derivation of the firstsequence from the second. The identity is determined by means of thecomputer program GAP provided in the GCG program package. Thus, GapGCGv8 is used with the following default parameters: GAP creationpenalty of 3.0 and GAP extension penalty of 0.1, the default scoringmatrix, for protein sequences. GAP uses the method ofNeedleman/Wunsch/Sellers to make alignments.

Detergent Compositions Comprising Other Enzymes

Various types of enzymes are very commonly included in detergentformulations for laundry cleaning and for hard surface cleaning. Onepurpose of these enzymes is to degrade soil binding substances. Thesesubstances bind or attract soil and stains onto the fabric or hardsurface. The degraded substances, and the associated soil and stain, arethen more easily removed during the washing process. Redeposition ofsoil, which can be increased by soil binding substances, is prevented.

The substance on which an enzyme acts is called the substrate. Enzymesare known to be “substrate specific”, i.e. each class of enzyme can onlydegrade one class of substances. For example, a protease can degradeproteins but cannot degrade starch. An amylase can degrade starch butcannot degrade proteins.

Because the soils and stains that are important for detergentformulators can contain many kinds of soil binding substances, a rangeof different enzymes, all with different substrate specificities havebeen developed for use in detergents. These include enzymes such asprotease, amylase, beta-glucanase, lipase, hemi-cellulase, cutinase,pectinase and pectate lyase.

Surprisingly it has been found that the endo-glucanase of the inventionare not substrate specific, in that they can provide detergency benefitson a wide range of soils and stains when used in combination with otherenzymes.

The endo-glucanase of the present invention includes, in addition to theenzymes specified by reference to SEQ ID NO 2, other endo-glucanaseswith surprisingly high a ntiredeposition effect. Two tests are providedin order to describe and identify these enzymes: 1) a test forendo-glucanase activity, and 2) a test for anti-redeposition effect.Enzymes which provide greater than the specified minimum performance onboth these tests are regarded, for the purpose of this invention, asendo-glucanases having anti-redeposition effect.

Thus in a preferred embodiment an endo-glucanase of the invention isused together with an amylase to provide improved detergency performanceon soils that contain starch. Such amylases comprise e.g. α- orβ-amylases of bacterial or fungal origin. Chemically or geneticallymodified mutants of such amylases are included in this connection.Relevant α-amylases include, for example, α-amylases obtainable fromBacillus species, in particular a special strain of B. licheniformis,described in more detail in GB 1296839. Relevant commercially availableamylases include Natalase™, Stainzyme®, Duramy®, Termamyl®, Termamyl™Ultra, Fungamyl® and BAN® (all available from Novozymes A/S, Bagsvaerd,Denmark), and Rapidasem and Maxamyl™ P (available from DSM, Holland).

In a preferred embodiment the alpha-amylase is derived from Bacillus sp.strains NCIB 12289, NCIB 12512, NCIB 12513 and DSM 9375. Especiallypreferred are the alphaamylases shown in SEQ ID NOS 1 and 2 of WO95/26397.

In another preferred embodiment the alpha-amylase is the AA560alpha-amylase derived from Bacillus sp. DSM 12649 disclosed as SEQ IDNO: 2 in WO 00/60060 (hereby incorporated by reference). Especiallypreferred are variants of the AA560 alpha-amylase, including the M560variant disclosed in Example 7 and 8 (hereby incorporated by reference).

Other useful amylases are CGTases (cyclodextrin glucanotransferases, EC2.4.1.19), e.g. those obtainable from species of Bacillus,Thermoanaerobactor or Themoanaerobacterium.

In another preferred embodiment an endo-glucanase of the invention isused together with a protease to provide improved detergency performanceon soils that contain protein. Such proteases comprise those of animal,vegetable or microbial origin. Proteases of microbial origin arepreferred. Chemically or genetically modified mutants of such proteasesare included in this connection. The protease may be a serine protease,preferably an alkaline microbial protease or a trypsin-like protease.Examples of alkaline proteases are subtilisins, especially those derivedfrom Bacillus, e.g., subtilisin Novo, subtilisin Carlsberg, subtilisin309, subtilisin 147 and subtilisin 168 (described in WO 89/06279).Examples of trypsin-like proteases are trypsin (e.g. of porcine orbovine origin) and the Fusarium protease described in WO 89/06270.

Relevant commercially available protease enzymes include Primase®,Durazym®, Everlase®, Kannase®, Alcalase®, Savinase® and Esperase® (allavailable from Novozymes A/S, Bagsvaerd, Denmark), Maxatase™, Maxaca™,Maxapem™ and Properase™ (available from DSM, Holland), Purafect™ andPurafect™ OXP (available from Genencor International, USA), andOpticlean™ and Optimase™ (available from by Solvay Enzymes).

In another preferred embodiment an endo-glucanase of the invention isused together with a lipase to provide improved detergency performanceon soils that contain fat or oil. Such lipases comprise those ofbacterial or fungal origin. Chemically or genetically modified mutantsof such lipases are included in this connection.

Examples of useful lipases include a Humicola lanuginosa lipase, e.g. asdescribed in EP 258 068 and EP 305 216; a Rhizomucor miehei lipase, e.g.as described in EP 238 023; a Candida lipase, such as a C. antarcticalipase, e.g. the C. antarctica lipase A or B described in EP 214 761; aPseudomonas lipase, such as one of those described in EP 721 981 (e.g. alipase obtainable from a Pseudomonas sp. SD705 strain having depositaccession number FERM BP-4772), in PCT/JP96/00426, in PCT/JP96/00454(e.g. a P. solanacearum lipase), in EP 571 982 or in WO 95/14783 (e.g. aP. mendocina lipase), a P. alcaligenes or P. pseudoalcaligenes lipase,e.g. as described in EP 218 272, a P. cepacia lipase, e.g. as describedin EP 331 376, a P. stutzeri lipase, e.g. as disclosed in GB 1,372,034,or a P. fluorescens lipase; a Bacillus lipase, e.g. a e.g. a B. subtilislipase (Dartois et al. (1993) Biochemica et Biophysica Acta1131:253-260), a B. stearothermophilus lipase (JP 64/744992) and a B.pumilus lipase (“O 91/16422).

Other potentially useful types of lipolytic enzymes include cutinases,e.g. a cutinase derived from Pseudomonas mendocina as described in WO88/09367, or a cutinase derived from Fusarium solani f. pisi (described,e.g., in WO 90/09446).

Suitable commercially available lipases include Lipex®, Lipolase® andLipolase Ultra® (available from Novozymes A/S), M1 Lipasem and Lipomax™(available from Genencor Inc.) and Lipase P “Amano” (available fromAmano Pharmaceutical Co. Ltd.). Commercially available cutinases includeLumafast™ from Genencor Inc.

In another preferred embodiment an endo-glucanase of the invention isused together with a hemi-cellulase to provide improved detergencyperformance on soils that contain hemicellulose and similarpolysaccharides. Such hemi-cellulases include xylanases, xyloglucanases,arabinofuranosidases, acetyl xylan esterases, glucuronidases, ferulicacid esterases, coumaric acid esterases, endo-galactanases, mannanases,endo- or exoarabinanases, exo-galactanases. Suitable mannanases includethose of bacterial or fungal origin. Chemically or genetically modifiedmutants are included.

In a preferred embodiment the mannanase is derived from a strain of thegenus Bacillus, especially Bacillus sp. 1633 disclosed in positions31-330 of SEQ ID NO:2 or in SEQ ID NO: 5 of WO 99/64619 or Bacillusagaradhaerens, for example from the type strain DSM 8721. A suitablemannanase is Mannaway® produced by Novozymes A/S.

Thus in a preferred embodiment an endo-glucanase of the invention isused together with a beta-glucanase (EC 3.2.1.6) to provide improveddetergency performance on soils that contain beta-glucans. Furtherpreferred beta-glucanases include lichenases and laminarinases.

Thus in a preferred embodiment an endo-glucanase of the invention isused together with pectinolytic enzymes such as a protopectinase,pectinase, polygalacturonase or pectate lyase to provide improveddetergency performance on pectinaceous soils. Suitable pectinolyticenzymes include those described in WO 99/27083, WO 99/27084, WO 00/55309and WO 02/092741. Suitable pectate lyases include those of bacterial orfungal origin. Chemically or genetically modified mutants are included.

In a preferred embodiment the pectate lyase is derived from a strain ofthe genus Bacillus, especially a strain of Bacillus substilis,especially Bacillus subtilis DSM14218 disclosed in SEQ ID NO:2 or avariant thereof disclosed in Example 6 of WO 02/092741.

Additionally, in a preferred embodiment an endo-glucanase of theinvention is used together with other enzymes of the classes: pectatelyase (EC 4.2.2.2), pectin lyase (EC 4.2.2.10), rhamnogalacturonan lyase(EC not defined), endo-1,4-galactanase (EC 3.2.1.89), xyloglucanase (ECnot defined), xylanase (EC 3.2.1.8), arabinanase (EC 3.2.1.99),alpha-L-arabinofuranosidase (EC 3.2.1.55), Mannan endo-1,4-mannosidase(EC 3.2.1.78), beta-mannosidase (EC 3.2.1.25), beta-1,3-1,4-glucanase(EC 3.2.1.73), rhamnogalacturonan hydrolase, exo-polygalacturonase (EC3.2.1.67), rhamnogalacturonase (EC not defined), Glucan1,3-beta-glucosidase (EC 3.2.1.58), Glucan endo-1,6-beta-glucosidase (EC3.2.1.75), Mannan endo-1,4-beta-mannosidase (EC 3.2.1.78),Endo-1,4-beta-xylanase (EC 3.2.1.8), Cellulose 1,4-cellobiosidase (EC3.2.1.91), cellobiohydrolase (EC 3.2.1.91). Polygalacturonases (EC3.2.1.15). Acetyl and methyl esterase enzymes such as:rhamnogalacturonan methyl esterase, rhamnogalacturonan acetyl esterase,pectin methylesterase (EC 3.1.1.11), pectin acetylesterase (EC notdefined), xylan methyl esterase, acetyl xylan esterase (EC 3.1.1.72),feruloyl esterase (EC 3.1.1.73), cinnamoyl esterase (EC 3.1.1.73) toprovide improved detergency performance on corresponding soils.

Detergent Composition of the Invention

The detergent compositions according to the present invention comprise asurfactant system, wherein the surfactant can be selected from nonionicand/or anionic and/or cationic and/or amphoteric and/or zwitterionicand/or semi-polar surfactants.

The surfactant is typically present at a level from 0.1% to 60% byweight.

The surfactant is preferably formulated to be compatible with enzymecomponents present in the composition. In liquid or gel compositions thesurfactant is most preferably formulated in such a way that it promotes,or at least does not degrade, the stability of any enzyme in thesecompositions.

Preferred systems to be used according to the present invention compriseas a surfactant one or more of the nonionic and/or anionic surfactantsdescribed herein.

Polyethylene, polypropylene, and polybutylene oxide condensates of alkylphenols are suitable for use as the nonionic surfactant of thesurfactant systems of the present invention, with the polyethylene oxidecondensates being preferred. These compounds include the condensationproducts of alkyl phenols having an alkyl group containing from about 6to about 14 carbon atoms, preferably from about 8 to about 14 carbonatoms, in either a straight chain or branched-chain configuration withthe alkylene oxide. In a preferred embodiment, the ethylene oxide ispresent in an amount equal to from about 2 to about 25 moles, morepreferably from about 3 to about 15 moles, of ethylene oxide per mole ofalkyl phenol. Commercially available nonionic surfactants of this typeinclude Igepal™ CO-630, marketed by the GAF Corporation; and Triton™X45, X-114, X-100 and X-102, all marketed by the Rohm & Haas Company.These surfactants are commonly referred to as alkylphenol alkoxylates(e.g., alkyl phenol ethoxylates).

The condensation products of primary and secondary aliphatic alcoholswith about 1 to about 25 moles of ethylene oxide are suitable for use asthe nonionic surfactant of the nonionic surfactant systems of thepresent invention. The alkyl chain of the aliphatic alcohol can eitherbe straight or branched, primary or secondary, and generally containsfrom about 8 to about 22 carbon atoms. Preferred are the condensationproducts of alcohols having an alkyl group containing from about 8 toabout 20 carbon atoms, more preferably from about 10 to about 18 carbonatoms, with from about 2 to about 10 moles of ethylene oxide per mole ofalcohol. About 2 to about 7 moles of ethylene oxide and most preferablyfrom 2 to 5 moles of ethylene oxide per mole of alcohol are present insaid condensation products. Examples of commercially available nonionicsurfactants of this type include Tergitol™ 15-S-9 (The condensationproduct of C₁₁-C₁₅ linear alcohol with 9 moles ethylene oxide),Tergitol™ 24-L-6 NMW (the condensation product of C₁₂-C₁₄ primaryalcohol with 6 moles ethylene oxide with a narrow molecular weightdistribution), both marketed by Union Carbide Corporation; Neodol™ 45-9(the condensation product of C₁₄-C₁₅ linear alcohol with 9 moles ofethylene oxide), Neodol™ 23-3 (the condensation product of C₁₂-C₁₃linear alcohol with 3.0 moles of ethylene oxide), Neodol™ 45-7 (thecondensation product of C₁₄-C₁₅ linear alcohol with 7 moles of ethyleneoxide), Neodol™ 45-5 (the condensation product of C₁₄-C₁₅ linear alcoholwith 5 moles of ethylene oxide) marketed by Shell Chemical Company,Kyro™ EOB (the condensation product of C₁₃-C₁₅ alcohol with 9 molesethylene oxide), marketed by The Procter & Gamble Company, and GenapolLA 050 (the condensation product of C₁₂-C₁₄ alcohol with 5 moles ofethylene oxide) marketed by Hoechst. Preferred range of HLB in theseproducts is from 8-11 and most preferred from 8-10.

Also useful as the nonionic surfactant of the surfactant systems of thepresent invention are alkylpolysaccharides disclosed in U.S. Pat. No.4,565,647, having a hydrophobic group containing from about 6 to about30 carbon atoms, preferably from about 10 to about 16 carbon atoms and apolysaccharide, e.g. a polyglycoside, hydrophilic group containing fromabout 1.3 to about 10, preferably from about 1.3 to about 3, mostpreferably from about 1.3 to about 2.7 saccharide units. Any reducingsaccharide containing 5 or 6 carbon atoms can be used, e.g., glucose,galactose and galactosyl moieties can be substituted for the glucosylmoieties (optionally the hydrophobic group is attached at the 2-, 3-,4-, etc. positions thus giving a glucose or galactose as opposed to aglucoside or galactoside). The intersaccharide bonds can be, e.g.,between the one position of the additional saccharide units and the 2-,3-, 4-, and/or 6-positions on the preceding saccharide units.

The preferred alkylpolyglycosides have the formulaR²O(C_(n)H_(2n)O)_(t)(glycosyl)_(x)wherein R² is selected from the group consisting of alkyl, alkylphenyl,hydroxyalkyl, hydroxyalkylphenyl, and mixtures thereof in which thealkyl groups contain from about 10 to about 18, preferably from about 12to about 14, carbon atoms; n is 2 or 3, preferably 2; t is from 0 toabout 10, preferably 0; and x is from about 1.3 to about 10, preferablyfrom about 1.3 to about 3, most preferably from about 1.3 to about 2.7.The glycosyl is preferably derived from glucose. To prepare thesecompounds, the alcohol or alkylpolyethoxy alcohol is formed first andthen reacted with glucose, or a source of glucose, to form the glucoside(attachment at the 1-position). The additional glycosyl units can thenbe attached between their 1-position and the preceding glycosyl units2-, 3-, 4, and/or 6-position, preferably predominantly the 2-position.

The condensation products of ethylene oxide with a hydrophobic baseformed by the condensation of propylene oxide with propylene glycol arealso suitable for use as the additional nonionic surfactant systems ofthe present invention. The hydrophobic portion of these compounds willpreferably have a molecular weight from about 1500 to about 1800 andwill exhibit water insolubility. The addition of polyoxyethylenemoieties to this hydrophobic portion tends to increase the watersolubility of the molecule as a whole, and the liquid character of theproduct is retained up to the point where the polyoxyethylene content isabout 50% of the total weight of the condensation product, whichcorresponds to condensation with up to about 40 moles of ethylene oxide.Examples of compounds of this type include certain of the commerciallyavailable Pluronic™ surfactants, marketed by BASF.

Also suitable for use as the nonionic surfactant of the nonionicsurfactant system of the present invention, are the condensationproducts of ethylene oxide with the product resulting from the reactionof propylene oxide and ethylenediamine. The hydrophobic moiety of theseproducts consists of the reaction product of ethylenediamine and excesspropylene oxide, and generally has a molecular weight of from about 2500to about 3000. This hydrophobic moiety is condensed with ethylene oxideto the extent that the condensation product contains from about 40% toabout 80% by weight of polyoxyethylene and has a molecular weight offrom about 5,000 to about 11,000. Examples of this type of nonionicsurfactant include certain of the commercially available Tetronic™compounds, marketed by BASF.

Preferred for use as the nonionic surfactant of the surfactant systemsof the present invention are polyethylene oxide condensates of alkylphenols, condensation products of primary and secondary aliphaticalcohols with from about 1 to about 25 moles of ethyleneoxide,alkylpolysaccharides, and mixtures hereof. Most preferred are C₈-C₁₄alkyl phenol ethoxylates having from 3 to 15 ethoxy groups and C₈-C₁₈alcohol ethoxylates (preferably C₁₀ avg.) having from 2 to 10 ethoxygroups, and mixtures thereof. Highly preferred nonionic surfactants arepolyhydroxy fatty acid amide surfactants of the formula

wherein R¹ is H, or R¹ is C₁₋₄ hydrocarbyl, 2-hydroxyethyl,2-hydroxypropyl or a mixture thereof, R² is C₅₋₃₁ hydrocarbyl, and Z isa polyhydroxyhydrocarbyl having a linear hydrocarbyl chain with at least3 hydroxyls directly connected to the chain, or an alkoxylatedderivative thereof. Preferably, R¹ is methyl, R² is straight C₁₁₋₁₅alkyl or C₁₆₋₁₈ alkyl or alkenyl chain such as coconut alkyl or mixturesthereof, and Z is derived from a reducing sugar such as glucose,fructose, maltose or lactose, in a reductive amination reaction.

Highly preferred anionic surfactants include alkyl alkoxylated sulfatesurfactants. Examples hereof are water soluble salts or acids of theformula RO(A)_(m)SO3M wherein R is an unsubstituted C₁₀-C-₂₄ alkyl orhydroxyalkyl group having a C₁₀-C₂₄ alkyl component, preferably aC₁₂-C₂₀ alkyl or hydro-xyalkyl, more preferably C₁₂-C₁₈ alkyl orhydroxyalkyl, A is an ethoxy or propoxy unit, m is greater than zero,typically between about 0.5 and about 6, more preferably between about0.5 and about 3, and M is H or a cation which can be, for example, ametal cation (e.g., sodium, potassium, lithium, calcium, magnesium,etc.), ammonium or substituted-ammonium cation. Alkyl ethoxylatedsulfates as well as alkyl propoxylated sulfates are contemplated herein.Specific examples of substituted ammonium cations include methyl-,dimethyl, trimethyl-ammonium cations and quaternary ammonium cationssuch as tetramethyl-ammonium and dimethyl piperdinium cations and thosederived from alkylamines such as ethylamine, diethylamine,triethylamine, mixtures thereof, and the like. Exemplary surfactants areC₁₂-C₁₈ alkyl polyethoxylate (1.0) sulfate (C₁₂-C₁₈E(1.0)M), C₁₂-C₁₈alkyl polyethoxylate (2.25) sulfate (C₁₂-C₁₈(2.25)M, and C₁₂-C₁₈ alkylpolyethoxylate (3.0) sulfate (C₁₂-C₁₈E(3.0)M), and C₁₂-C₁₈ alkylpolyethoxylate (4.0) sulfate (C₁₂-C₁₈E(4.0)M), wherein M is convenientlyselected from sodium and potassium.

Suitable anionic surfactants to be used are alkyl ester sulfonatesurfactants including linear esters of C₈-C₂₀ carboxylic acids (i.e.,fatty acids) which are sulfonated with gaseous SO₃ according to “TheJournal of the American Oil Chemists Society”, 52 (1975), pp. 323-329.Suitable starting materials would include natural fatty substances asderived from tallow, palm oil, etc.

The preferred alkyl ester sulfonate surfactant, especially for laundryapplications, comprises alkyl ester sulfonate surfactants of thestructural formula:

wherein R³ is a C₈-C₂₀ hydrocarbyl, preferably an alkyl, or combinationthereof, R⁴ is a C₁-C₈ hydrocarbyl, preferably an alkyl, or combinationthereof, and M is a cation which forms a water soluble salt with thealkyl ester sulfonate. Suitable salt-forming cations include metals suchas sodium, potassium, and lithium, and substituted or unsubstitutedammonium cations, such as monoethanolamine, diethonolamine, andtriethanolamine. Preferably, R³ is C₁₀-C₁₆ alkyl, and R⁴ is methyl,ethyl or isopropyl. Especially preferred are the methyl ester sulfonateswherein R³ is C₁₀-C₁₆ alkyl.

Other suitable anionic surfactants include the alkyl sulfate surfactantswhich are water soluble salts or acids of the formula ROSO₃M wherein Rpreferably is a C₁₀-C₂₄ hydrocarbyl, preferably an alkyl or hydroxyalkylhaving a C₁₀-C₂₀ alkyl component, more preferably a C₁₂-C₁₈ alkyl orhydroxyalkyl, and M is H or a cation, e.g., an alkali metal cation (e.g.sodium, potassium, lithium), or ammonium or substituted ammonium (e.g.methyl-, dimethyl-, and trimethyl ammonium cations and quaternaryammonium cations such as tetramethyl-ammonium and dimethyl piperdiniumcations and quaternary ammonium cations derived from alkylamines such asethylamine, diethylamine, triethylamine, and mixtures thereof, and thelike). Typically, alkyl chains of C₁₂-C₁₆ are preferred for lower washtemperatures (e.g. below about 50° C.) and C₁₆-C₁₈ alkyl chains arepreferred for higher wash temperatures (e.g. above about 50° C.).

Other anionic surfactants useful for detersive purposes can also beincluded in the laundry detergent compositions of the present invention.Theses can include salts (including, for example, sodium, potassium,ammonium, and substituted ammonium salts such as mono- di- andtriethanolamine salts) of soap, C₈-C₂₂ primary or secondaryalkanesulfonates, C₈-C₂₄ olefinsulfonates, sulfonated polycarboxylicacids prepared by sulfonation of the pyrolyzed product of alkaline earthmetal citrates, e.g., as described in British patent specification No.1,082,179, C₈-C₂₄ alkylpolyglycolethersulfates (containing up to 10moles of ethylene oxide); alkyl glycerol sulfonates, fatty acyl glycerolsulfonates, fatty oleyl glycerol sulfates, alkyl phenol ethylene oxideether sulfates, paraffin sulfonates, alkyl phosphates, isethionates suchas the acyl isethionates, N-acyl taurates, alkyl succinamates andsulfosuccinates, monoesters of sulfosuccinates (especially saturated andunsaturated C₁₂-C₁₈ monoesters) and diesters of sulfosuccinates(especially saturated and unsaturated C₆-C₁₂ diesters), acylsarcosinates, sulfates of alkylpolysaccharides such as the sulfates ofalkylpolyglucoside (the nonionic nonsulfated compounds being describedbelow), branched primary alkyl sulfates, and alkyl polyethoxycarboxylates such as those of the formula RO(CH₂CH₂O)_(k)—CH₂C00-M+wherein R is a C₈-C₂₂ alkyl, k is an integer from 1 to 10, and M is asoluble salt forming cation. Resin acids and hydrogenated resin acidsare also suitable, such as rosin, hydrogenated rosin, and resin acidsand hydrogenated resin acids present in or derived from tall oil.

Alkylbenzene sulfonates are highly preferred. Especially preferred arelinear (straight-chain) alkyl benzene sulfonates (LAS) wherein the alkylgroup preferably contains from 10 to 18 carbon atoms.

Further examples are described in “Surface Active Agents and Detergents”(Vol. 1 and 11 by Schwartz, Perrry and Berch). A variety of suchsurfactants are also generally disclosed in U.S. Pat. No. 3,929,678,(Column 23, line 58 through Column 29, line 23, herein incorporated byreference).

When included therein, the detergent compositions of the presentinvention typically comprise from about 1% to about 40%, preferably fromabout 3% to about 25% by weight of such anionic surfactants.

The laundry detergent compositions of the present invention may alsocontain cationic, amphoteric, zwitterionic, and semi-polar surfactants,as well as the nonionic and/or anionic surfactants other than thosealready described herein.

Cationic detersive surfactants suitable for use in the laundry detergentcompositions of the present invention are those having one long-chainhydrocarbyl group. Examples of such cationic surfactants include theammonium surfactants such as alkyltrimethylammonium halogenides, andthose surfactants having the formula:[R²(OR³)_(y)][R⁴(OR³)_(y)]₂R⁵N⁺X⁻wherein R² is an alkyl or alkyl benzyl group having from about 8 toabout 18 carbon atoms in the alkyl chain, each R³ is selected form thegroup consisting of —CH₂CH₂—, —CH₂CH(CH₃)—, —CH₂CH(CH₂OH)—, —CH₂CH₂CH₂—,and mixtures thereof; each R⁴ is selected from the group consisting ofC₁-C₄ alkyl, C₁-C₄ hydroxyalkyl, benzyl ring structures formed byjoining the two R⁴ groups, —CH₂CHOHCHOHCOR⁶CHOHCH₂OH, wherein R⁶ is anyhexose or hexose polymer having a molecular weight less than about 1000,and hydrogen when y is not 0; R⁵ is the same as R⁴ or is an alkyl chain,wherein the total number of carbon atoms of R² plus R⁵ is not more thanabout 18; each y is from 0 to about 10, and the sum of the y values isfrom 0 to about 15; and X is any compatible anion.

Highly preferred cationic surfactants are the water soluble quaternaryammonium compounds useful in the present composition having the formula:R₁R₂R₃R₄N⁺X⁻  (i)wherein R₁ is C₈-C₁₆ alkyl, each of R₂, R₃ and R₄ is independently C₁-C₄alkyl, C₁-C₄ hydroxy alkyl, benzyl, and —(C₂H₄₀)_(x) H where x has avalue from 2 to 5, and X is an anion. Not more than one of R₂, R₃ or R₄should be benzyl.

The preferred alkyl chain length for R₁ is C₁₂-C₁₅, particularly wherethe alkyl group is a mixture of chain lengths derived from coconut orpalm kernel fat or is derived synthetically by olefin build up or OXOalcohols synthesis.

Preferred groups for R₂, R₃ and R₄ are methyl and hydroxyethyl groupsand the anion X may be selected from halide, methosulphate, acetate andphosphate ions.

Examples of suitable quaternary ammonium compounds of formulae (i) foruse herein are:

-   -   coconut trimethyl ammonium chloride or bromide;    -   coconut methyl dihydroxyethyl ammonium chloride or bromide;    -   decyl triethyl ammonium chloride;    -   decyl dimethyl hydroxyethyl ammonium chloride or bromide;    -   C₁₂₋₁₅ dimethyl hydroxyethyl ammonium chloride or bromide;    -   coconut dimethyl hydroxyethyl ammonium chloride or bromide;    -   myristyl trimethyl ammonium methyl sulphate;    -   lauryl dimethyl benzyl ammonium chloride or bromide;    -   lauryl dimethyl (ethenoxy)₄ ammonium chloride or bromide;    -   choline esters (compounds of formula (i) wherein R₁ is        di-alkyl imidazolines [compounds of formula (i)].

Other cationic surfactants useful herein are also described in U.S. Pat.No. 4,228,044 and in EP 000 224.

When included therein, the detergent compositions of the presentinvention typically comprise from 0.2% to about 25%, preferably fromabout 1% to about 8% by weight of such cationic surfactants.

Amphoteric surfactants are also suitable for use in the detergentcompositions of the present invention. These surfactants can be broadlydescribed as aliphatic derivatives of secondary or tertiary amines, or aliphatic derivatives of heterocyclic secondary and tertiary amines inwhich the aliphatic radical can be straight- or branched-chain. One ofthe aliphatic substituents contains at least about 8 carbon atoms,typically from about 8 to about 18 carbon atoms, and at least onecontains an anionic water-solubilizing group, e.g. carboxy, sulfonate,sulfate. See U.S. Pat. No. 3,929,678 (column 19, lines 18-35) forexamples of amphoteric surfactants.

When included therein, the detergent compositions of the presentinvention typically comprise from 0.2% to about 15%, preferably fromabout 1% to about 10% by weight of such amphoteric surfactants.

Zwitterionic surfactants a re also suitable for use in detergentcompositions. These surfactants can be broadly described as derivativesof secondary and tertiary amines, derivatives of heterocyclic secondaryand tertiary amines, or derivatives of quaternary ammonium, quaternaryphosphonium or tertiary sulfonium compounds. See U.S. Pat. No. 3,929,678(column 19, line 38 through column 22, line 48) for examples ofzwitterionic surfactants.

When included therein, the detergent compositions of the presentinvention typically comprise from 0.2% to about 15%, preferably fromabout 1% to about 10% by weight of such zwitterionic surfactants.

Semi-polar nonionic surfactants are a special category of nonionicsurfactants which include water-soluble amine oxides containing onealkyl moiety of from about 10 to about 18 carbon atoms and 2 moietiesselected from the group consisting of alkyl groups and hydroxyalkylgroups containing from about 1 to about 3 carbon atoms; water solublephosphine oxides containing one alkyl moiety of from about 10 to about18 carbon atoms and 2 moieties selected from the group consisting ofalkyl groups and hydroxyalkyl groups containing from about 1 to about 3carbon atoms; and water-soluble sulfoxides containing one alkyl moietyfrom about 10 to about 18 carbon atoms and a moiety selected from thegroup consisting of alkyl and hydroxyalkyl moieties of from about 1 toabout 3 carbon atoms.

Semi-polar nonionic detergent surfactants include the amine oxidesurfactants having the formula:

wherein R³ is an alkyl, hydroxyalkyl, or alkyl phenyl group or mixturesthereof containing from about 8 to about 22 carbon atoms; R⁴ is analkylene or hydroxyalkylene group containing from about 2 to about 3carbon atoms or mixtures thereof; x is from 0 to about 3: and each R⁵ isan alkyl or hydroxyalkyl group containing from about 1 to about 3 carbonatoms or a polyethylene oxide group containing from about 1 to about 3ethylene oxide groups. The R⁵ groups can be attached to each other,e.g., through an oxygen or nitrogen atom, to form a ring structure.

These amine oxide surfactants in particular include C₁₀-C₁₈ alkyldimethyl amine oxides and C₈-C₁₂ alkoxy ethyl dihydroxy ethyl amineoxides.

When included therein, the detergent compositions of the presentinvention typically comprise from 0.2% to about 15%, preferably fromabout 1% to about 10% by weight of such semi-polar nonionic surfactants.

Builder System

The compositions according to the present invention may further comprisea builder system. Any conventional builder system is suitable for useherein including aluminosilicate materials, silicates, polycarboxylatesand fatty acids, materials such as ethylenediamine tetraacetate, metalion sequestrants such as aminopolyphosphonates, particularlyethylenediamine tetramethylene phosphonic acid and diethylene triaminepentamethylenephosphonic acid. Though less preferred for obviousenvironmental reasons, phosphate builders can also be used herein.

Suitable builders can be an inorganic ion exchange material, commonly aninorganic hydrated aluminosilicate material, more particularly ahydrated synthetic zeolite such as hydrated zeolite A, X, B, HS or MAP.

Another suitable inorganic builder material is layered silicate, e.g.SKS-6 (Hoechst). SKS-6 is a crystalline layered silicate consisting ofsodium silicate (Na₂Si₂O₅).

Suitable polycarboxylates containing one carboxy group include lacticacid, glycolic acid and ether derivatives thereof as disclosed inBelgian Patent Nos. 831, 368, 821, 369 and 821,370. Polycarboxylatescontaining two carboxy groups include the water-soluble salts ofsuccinic acid, malonic acid, (ethylenedioxy) diacetic acid, maleic acid,diglycollic acid, tartaric acid, tartronic acid and fumaric acid, aswell as the ether carboxylates described in DE 2,446,686, and 2,446,487,U.S. Pat. No. 3,935,257 and the sulfinyl carboxylates described inBelgian Patent No. 840,623. Polycarboxylates containing three carboxygroups include, in particular, water-soluble citrates, aconitrates andcitraconates as well as succinate derivatives such as thecarboxymethyloxysuccinates described in British Patent No. 1,379,241,lactoxysuccinates described in Netherlands Application 7205873, and theoxypolycarboxylate materials such as 2-oxa-1,1,3-propane tricarboxylatesdescribed in British Patent No. 1,387,447.

Polycarboxylates containing four carboxy groups include oxydisuccinatesdisclosed in British Patent No. 1,261,829, 1,1,2,2,-ethanetetracarboxylates, 1,1,3,3-propane tetracarboxylates containing sulfosubstituents include the sulfosuccinate derivatives disclosed in BritishPatent Nos. 1,398,421 and 1,398,422 and in U.S. Pat. No. 3,936,448, andthe sulfonated pyrolysed citrates described in British Patent No.1,082,179, while polycarboxylates containing phosphone substituents aredisclosed in British Patent No. 1,439,000.

Alicyclic and heterocyclic polycarboxylates includecyclopentane-cis,cis-cistetracarboxylates, cyclopentadienidepentacarboxylates, 2,3,4,5-tetrahydro-furan-cis, cis,cistetracarboxylates, 2,5-tetrahydro-furan-cis, discarboxylates,2,2,5,5,-tetrahydrofuran tetracarboxylates,1,2,3,4,5,6-hexane-hexacarboxylates and carboxymethyl derivatives ofpolyhydric alcohols such as sorbitol, mannitol and xylitol. Aromaticpolycarboxylates include mellitic acid, pyromellitic acid and thephthalic acid derivatives disclosed in British Patent No. 1,425,343.

Of the above, the preferred polycarboxylates are hydroxy-carboxylatescontaining up to three carboxy groups per molecule, more particularlycitrates.

Preferred builder systems for use in the present compositions include amixture of a water-insoluble aluminosilicate builder such as zeolite Aor of a layered silicate (SKS-6), and a water-soluble carboxylatechelating agent such as citric acid.

A suitable chelant for inclusion in the detergent compositions inaccordance with the invention is ethylenediamine-N,N′-disuccinic acid(EDDS) or the alkali metal, alkaline earth metal, ammonium, orsubstituted ammonium salts thereof, or mixtures thereof. Preferred EDDScompounds are the free acid form and the sodium or magnesium saltthereof. Examples of such preferred sodium salts of EDDS include Na₂EDDSand Na₄EDDS. Examples of such preferred magnesium salts of EDDS includeMgEDDS and Mg₂EDbS. The magnesium salts are the most preferred forinclusion in compositions in accordance with the invention.

Other builder materials that can form part of the builder system for usein granular compositions include inorganic materials such as alkalimetal carbonates, bicarbonates, silicates, and organic materials such asthe organic phosphonates, amino polyalkylene phosphonates and aminopolycarboxylates.

Other suitable water-soluble organic salts are the homo- or co-polymericacids or their salts, in which the polycarboxylic acid comprises atleast two carboxyl radicals separated form each other by not more thantwo carbon atoms.

Polymers of this type are disclosed in GB-A-1,596,756. Examples of suchsalts are polyacrylates of MW 2000-5000 and their copolymers with maleicanhydride, such copolymers having a molecular weight of from 20,000 to70,000, especially about 40,000.

Detergency builder salts are normally included in amounts of from 5% to80% by weight of the composition. Preferred levels of builder for liquiddetergents are from 5% to 30%.

Bleaching agents: Additional optional detergent ingredients that can beincluded in the detergent compositions of the present invention includebleaching agents such as perborate PB1, PB4 and percarbonate. Thesebleaching agent components can include one or more oxygen bleachingagents and, depending upon the bleaching agent chosen, one or morebleach activators. Present oxygen bleaching compounds will typically bepresent at levels of from about 1% to about 25%. In general, bleachingcompounds are optional added components in non-liquid formulations, e.g.granular detergents.

The bleaching agent component for use herein can be any of the bleachingagents useful for detergent compositions including oxygen bleaches aswell as others known in the art.

The bleaching agent suitable for the present invention can be anactivated or nonactivated bleaching agent.

One category of oxygen bleaching agent that can be used encompassespercarboxylic acid bleaching agents and salts thereof. Suitable examplesof this class of agents include magnesium monoperoxyphthalatehexahydrate, the magnesium salt of meta-chloro perbenzoic acid,4-nonylamino-4-oxoperoxybutyric acid and diperoxydodecanedioic acid.Such bleaching agents are disclosed in U.S. Pat. No. 4,483,781, EP 0 133354 and U.S. Pat. No. 4,412,934. Highly preferred bleaching agents alsoinclude 6-nonylamino-6-oxoperoxycaproic acid as described in U.S. Pat.No. 4,634,551.

Another category of bleaching agents encompasses the halogen bleachingagents. Examples of hypohalite bleaching agents, for example, includetrichloro isocyanuric acid and the sodium and potassiumdichloroisocyanurates and N-chloro and N-bromo alkane sulphonamides.Such materials are normally added at 0.5-10% by weight of the finishedproduct, preferably 1-5% by weight. Such halogen bleaching agents aregenerally less preferred for use in enzymatic detergents.

The hydrogen peroxide releasing agents can be used in combination withbleach activators such as tetra-acetylethylenediamine (TAED),nonanoyloxybenzenesulfonate (NOBS, described in U.S. Pat. No.4,412,934), 3,5-trimethyl-hexsanoloxybenzenesulfonate (ISONOBS,described in EP 120 591) or pentaacetylglucose (PAG), which areperhydrolyzed to form a peracid as the active bleaching species, leadingto improved bleaching effect. In addition, very suitable are the bleachactivators C8 (6-octanamido-caproyl) oxybenzene-sulfonate,C9(6-nonanamido caproyl) oxybenzenesulfonate and C10 (6-decanamidocaproyl) oxybenzenesulfonate or mixtures thereof. Also suitableactivators are acylated citrate esters such as disclosed in EuropeanPatent Application No. 91870207.7.

Useful bleaching agents, including peroxyacids and bleaching systemscomprising bleach activators and peroxygen bleaching compounds for usein cleaning compositions according to the invention are described inapplication U.S. Ser. No. 08/136,626.

The hydrogen peroxide may also be present by adding an enzymatic system(i.e. an enzyme and a substrate therefore) which is capable ofgeneration of hydrogen peroxide at the beginning or during the washingand/or rinsing process. Such enzymatic systems are disclosed in EuropeanPatent Application EP 0 537 381.

Bleaching agents other than oxygen bleaching agents are also known inthe art and can be utilized herein. One type of non-oxygen bleachingagent of particular interest includes photoactivated bleaching agentssuch as the sulfonated zinc and/or aluminium phthalocyanines. Thesematerials can be deposited upon the substrate during the washingprocess. Upon irradiation with light, in the presence of oxygen, such asby hanging clothes out to dry in the daylight, the sulfonated zincphthalocyanine is activated and, consequently, the substrate isbleached. Preferred zinc phthalocyanine and a photoactivated bleachingprocess are described in U.S. Pat. No. 4,033,718. Typically, detergentcomposition will contain about 0.025% to about 1.25%, by weight, ofsulfonated zinc phthalocyanine.

Bleaching agents may also comprise a manganese catalyst. The manganesecatalyst may, e.g., be one of the compounds described in “Efficientmanganese catalysts for low-temperature bleaching”, Nature (369) 1994,pp. 637-639.

Suds suppressors: Another optional ingredient is a suds suppressor,exemplified by silicones, and silica-silicone mixtures. Silicones cangenerally be represented by alkylated polysiloxane materials, whilesilica is normally used in finely divided forms exemplified by silicaaerogels and xerogels and hydrophobic silicas of various types. Thesesmaterials can be incorporated as particulates, in which the sudssuppressor is advantageously releasably incorporated in a water-solubleor water-dispersible, substantially non surface-active detergentimpermeable carrier. Alternatively the suds suppressor can be dissolvedor dispersed in a liquid carrier and applied by spraying onto one ormore of the other components.

A preferred silicone suds controlling agent is disclosed in U.S. Pat.No. 3,933,672. Other particularly useful suds suppressors are theself-emulsifying silicone suds suppressors, described in German PatentApplication DTOS 2,646,126. An example of such a compound is DC-544,commercially available form Dow Corning, which is a siloxane-glycolcopolymer. Especially preferred suds controlling agent are the sudssuppressor system comprising a mixture of silicone oils and2-alkyl-alkanols. Suitable 2-alkyl-alkanols are 2-butyl-octanols, whichare commercially available under the trade name Isofol 12 R.

Such suds suppressor system are described in European Patent ApplicationE P 0 593 841.

Especially preferred silicone suds controlling agents are described inEuropean Patent Application No. 92201649.8. Said compositions cancomprise a silicone/silica mixture in combination with fumed nonporoussilica such as Aerosil®.

The suds suppressors described above are normally employed at levels offrom 0.001% to 2% by weight of the composition, preferably from 0.01% to1% by weight.

Other components: Other components used in detergent compositions may beemployed such as soil-suspending or anti-redeposition agents,soil-releasing agents, optical brighteners, abrasives, bactericides,tarnish inhibitors, coloring agents, and/or encapsulated ornonencapsulated perfumes.

Especially suitable encapsulating materials are water soluble capsuleswhich consist of a matrix of polysaccharide and polyhydroxy compoundssuch as described in GB 1,464,616.

Other suitable water soluble encapsulating materials comprise dextrinsderived from ungelatinized starch acid esters of substituteddicarboxylic acids such as described in U.S. Pat. No. 3,455,838. Theseacid-ester dextrins are, preferably, prepared from such starches as waxymaize, waxy sorghum, sago, tapioca and potato. Suitable examples of saidencapsulation materials include N-Lok manufactured by National Starch.The N-Lok encapsulating material consists of a modified maize starch andglucose. The starch is modified by adding monofunctional substitutedgroups such as octenyl succinic acid anhydride.

Typical anti-redeposition agents used in detergents includewater-soluble, generally organic colloids, including for example thewater-soluble salts of polymeric carboxylic acids such as polyacrylicacid or polymaleic acid or co-polymers thereof, glue, gelatine, salts ofether carboxylic acids or ether sulfonic acids of starch or cellulose orsalts of sulfuric acid esters of cellulose or starch. Water-solublepolyamides containing acidic groups are also used as anti-redepositionagent. Soluble starch preparations and other starch products than thosementioned above, for example partly hydrolyzed starch, may also be used.Sodium carboxymethyl cellulose, methyl cellulose, hydroxyethylcellulose, methyl hydroxyethyl cellulose and mixtures thereof arepreferably used. These materials are normally used at levels of from0.05% to 10% by weight, more preferably form 0.2% to 8%, most preferablyfrom 0.5% to 6% by weight of the composition.

Preferred optical brighteners are anionic in character, examples ofwhich are disodium4,4′-bis-(2-diethanolamino-4-anilino-s-triazin-6-ylamino)stilbene-2:2′disulpho-nate, disodium4,4′-bis-(2-morpholino-4-anilino-s-triazin-6-ylamino-stilbene-2:2′-disulphonate,disodium4,4′-bis-(2,4-dianilino-s-triazin-6-ylamino)stilbene-2,2′-disulphonate,monosodium4′,4″-bis-(2,4-dianilino-s-tri-azin-6-ylamino)stilbene-2-sulphonate,disodium4,4′-bis-(2-anilino-4-(N-methyl-N-2-hydroxyethylamino)-s-triazin-6-ylamino)stilbene-2,2′-disulphonate, disodium4,4′-bis-(4-phenyl-2,1,3-triazol-2-yl)-stilbene-2,2′ disulphonate,disodium4,4′bis(2-anilino4-(1-methyl-2-hydroxyethylamino)-striazin-6-ylami-no)stilbene-2,2′disulphonate,sodium 2(stilbyl-4″-(naphtho-1′,2′:4,5)-1,2,3,-triazole-2″-sulphonateand 4,4′-bis(2-sulphostyryl)biphenyl.

Other useful polymeric materials are the polyethylene glycols,particularly those of molecular weight 1000-10000, more particularly2000 to 8000 and most preferably about 4000. These are used at levels offrom 0.20% to 5% more preferably from 0.25% to 2.5% by weight. Thesepolymers and the previously mentioned homo- or co-polymericpoly-carboxylate salts are valuable for improving whiteness maintenance,fabric ash deposition, and cleaning performance on clay, proteinaceousand oxidizable soils in the presence of transition metal impurities.

Soil release agents useful in compositions of the present invention areconventionally copolymers or terpolymers of terephthalic acid withethylene glycol and/or propylene glycol units in various arrangements.Examples of such polymers are disclosed in U.S. Pat. Nos. 4,116,885 and4,711,730 and EP 0 272 033. A particular preferred polymer in accordancewith EP 0 272 033 has the formula:(CH₃(PEG)₄₃)_(0.75)(POH)_(0.25)[T-PO)_(2.8)(T-PEG)_(0.4)]T(POH)_(0.25)((PEG)₄₃CH₃)_(0.75)where PEG is —(OC₂H₄)O—, PO is (OC₃H₆O) and T is (pOOC₆H₄CO).

Also very useful are modified polyesters as random copolymers ofdimethyl terephthalate, dimethyl sulfoisophthalate, ethylene glycol and1,2-propanediol, the end groups consisting primarily of sulphobenzoateand secondarily of mono esters of ethylene glycol and/or1,2-propanediol. The target is to obtain a polymer capped at both end bysulphobenzoate groups, “primarily”, in the present context most of saidcopolymers herein will be endcapped by sulphobenzoate groups. However,some copolymers will be less than fully capped, and therefore their endgroups may consist of monoester of ethylene glycol and/or1,2-propanediol, thereof consist “secondarily” of such species.

The selected polyesters herein contain about 46% by weight of dimethylterephthalic acid, about 16% by weight of 1,2-propanediol, about 10% byweight ethylene glycol, about 13% by weight of dimethyl sulfobenzoicacid and about 15% by weight of sulfoisophthalic acid, and have amolecular weight of about 3.000. The polyesters and their method ofpreparation are described in detail in EP 311 342.

Softening agents: Fabric softening agents can also be incorporated intolaundry detergent compositions in accordance with the present invention.These agents may be inorganic or organic in type. Inorganic softeningagents are exemplified by the smectite clays disclosed in GB-A-1 400898and in U.S. Pat. No. 5,019,292. Organic fabric softening agents includethe water insoluble tertiary amines as disclosed in GB-A1 514 276 and EP0 011 340 and their combination with mono C₁₂-C₁₄ quaternary ammoniumsalts are disclosed in EP-B-0 026 528 and di-long-chain amides asdisclosed in EP 0 242 919. Other useful organic ingredients of fabricsoftening systems include high molecular weight polyethylene oxidematerials as disclosed in EP 0 299 575 and 0 313 146.

Levels of smectite clay are normally in the range from 5% to 15%, morepreferably from 8% to 12% by weight, with the material being added as adry mixed component to the remainder of the formulation. Organic fabricsoftening agents such as the water-insoluble tertiary amines or di-longchain amide materials are incorporated at levels of from 0.5% to 5% byweight, normally from 1% to 3% by weight whilst the high molecularweight polyethylene oxide materials and the water soluble cationicmaterials are added at levels of from 0.1% to 2%, normally from 0.15% to1.5% by weight. These materials are normally added to the spray driedportion of the composition, although in some instances it may be moreconvenient to add them as a dry mixed particulate, or spray them asmolten liquid on to other solid components of the composition.

Polymeric dye-transfer inhibiting agents: The detergent compositionsaccording to the present invention may also comprise from 0.001% to 10%,preferably from 0.01% to 2%, more preferably form 0.05% to 1% by weightof polymeric dye-transfer inhibiting agents. Said polymeric dye-transferinhibiting agents are normally incorporated into detergent compositionsin order to inhibit the transfer of dyes from colored fabrics ontofabrics washed therewith. These polymers have the ability of complexingor adsorbing the fugitive dyes washed out of dyed fabrics before thedyes have the opportunity to become attached to other articles in thewash.

Especially suitable polymeric dye-transfer inhibiting agents arepolyamine N-oxide polymers, copolymers of N-vinyl-pyrrolidone andN-vinylimidazole, polyvinylpyrrolidone polymers, polyvinyloxazolidonesand polyvinylimidazoles or mixtures thereof.

Addition of such polymers also enhances the performance of the enzymesaccording the invention.

Enzymes

A detergent composition of the invention may in an embodiment of theinvention besides the endo-glucanase having anti-redeposition effect asdefined above, comprise other enzyme(s) which provides cleaningperformance and/or fabric care benefits.

Such enzymes include certain proteases, lipases, cutinases, cellulases,amylases, peroxidases, oxidases (e.g. laccases), and hemicellulases suchas mannanase and pectate lyase.

Proteases: Any protease suitable for use in alkaline solutions can beused. Suitable proteases include those of animal, vegetable or microbialorigin. Microbial origin is preferred. Chemically or geneticallymodified mutants are included. The protease may be a serine protease,preferably an alkaline microbial protease or a trypsin-like protease.Examples of alkaline proteases are subtilisins, especially those derivedfrom Bacillus, e.g., subtilisin Novo, subtilisin Carlsberg, subtilisin309, subtilisin 147 and subtilisin 168 (described in WO 89/06279).

Examples of trypsin-like proteases are trypsin (e.g. of porcine orbovine origin) and the Fusarium protease described in WO 89/06270.

Preferred commercially available protease enzymes include those soldunder the trade names Everlase™, Kannase™, Alcalase™, Savinase™,Primase™, Durazym™, and Esperase™ by Novozymes A/S (Denmark), those soldunder the tradename Maxatase, Maxacal, Maxapem, Properase, Purafect andPurafect OXP by Genenoor International, and those sold under thetradename Opticlean and Optimase by Solvay Enzymes. Protease enzymes maybe incorporated into the compositions in accordance with the inventionat a level of from 0.000001% to 2% of enzyme protein by weight of thecomposition, preferably at a level of from 0.00001% to 1% of enzymeprotein by weight of the composition, more preferably at a level of from0.0001% to 0.5% of enzyme protein by weight of the composition, evenmore preferably at a level of from 0.001% to 0.2% of enzyme protein byweight of the composition.

Lipases: Any lipase suitable for use in alkaline solutions can be used.Suitable lipases include those of bacterial or fungal origin. Chemicallyor genetically modified mutants are included.

Examples of useful lipases include a Humicola lanuginosa lipase, e.g.,as described in EP 258 068 and EP 305 216, a Rhizomucor miehei lipase,e.g., as described in EP 238 023, a Candida lipase, such as a C.antarctica lipase, e.g., the C. antarctica lipase A or B described in EP214 761, a Pseudomonas lipase such as a P. alcaligenes and P.pseudoalcaligenes lipase, e.g., as described in EP 218 272, a P. cepacialipase, e.g., as described in EP 331 376, a P. stutzeri lipase, e.g., asdisclosed in GB 1,372,034, a P. fluorescens lipase, a Bacillus lipase,e.g., a B. subtilis lipase (Dartois et al., (1993), Biochemica etBiophysica acta 1131, 253-260), a B. stearothermophilus lipase (JP64/744992) and a B. pumilus lipase (WO 91/16422).

Furthermore, a number of cloned lipases may be useful, including thePenicillium camembertii lipase described by Yamaguchi et al., (1991),Gene 103, 61-67), the Geotricum candidum lipase (Schimada, Y. et al.,(1989), J. Biochem., 106, 383-388), and various Rhizopus lipases such asa R. delemar lipase (Hass, M. J et al., (1991), Gene 109, 117-113), a R.niveus lipase (Kugimiya et al., (1992), Biosci. Biotech. Biochem.56,716-719) and a R. oryzae lipase.

Other types of lipolytic enzymes such as cutinases may also be useful,e.g., a cutinase derived from Pseudomonas mendocina as described in WO88/09367, or a cutinase derived from Fusarium solani pisi (e.g.described in WO 90/09446).

In a preferred embodiment the lipase is a variant of Humicola lanuginosaDSM 4109 as described in WO 00/60063. Especially preferred are thevariants disclosed in the Example in WO 00/60063 with improved firstwash performance., i.e., T231R+N233R;G91A+D96W+E99K+G263Q+L264A+1265T+G266D+T267A+L269N+270AGGFSWRRYRSAESVDKRATMTDAELEKKLNSYVQMDKEYVKNNQARS; R209P+T231 R+N233R; N33Q+D96S+T231 R+N233R+Q249R; E99N+N 101S+T231 R+N233R+Q249R; E99N+N101 S+T231 R+N233R+Q249R.

Especially suitable lipases are lipases such as M1 Lipase™ and Lipomax™(Genencor), Lipolase™ and Lipolase Ultra™, Lipex™ (Novozymes A/S), andLipase P “Amano” (Amano Pharmaceutical Co. Ltd.). Suitable cutinasesinclude Lumafast™ available from Genencor Inc.

The lipases are normally incorporated in the detergent composition at alevel of from 0.000001% to 2% of enzyme protein by weight of thecomposition, preferably at a level of from 0.00001% to 1% of enzymeprotein by weight of the composition, more preferably at a level of from0.0001% to 0.5% of enzyme protein by weight of the composition, evenmore preferably at a level of from 0.001% to 0.2% of enzyme protein byweight of the composition.

Amylases: Any amylase (alpha and/or beta) suitable for use in alkalinesolutions can be used. Suitable amylases include those of bacterial orfungal origin. Chemically or genetically modified mutants are included.Amylases include, for example, alpha-amylases obtained from a specialstrain of B. licheniformis, described in more detail in GB 1,296,839.Commercially available amylases are Natalase™, Termamyl™ Ultra,DuraMyl™, TermaMyl™, Fungamy™ and BAN™ (available from Novozymes A/S)and Rapidase™ and Maxamyl P™ (available from Genencor Inc.).

In a preferred embodiment the alpha-amylase is derived from Bacillus sp.strains NCIB 12289, NCIB 12512, NCIB 12513 and DSM 9375. Especiallypreferred are the alpha-amylases shown in SEQ ID NOS 1 and 2 of WO95/26397.

In another preferred embodiment the alpha-amylase is the AA560alpha-amylase derived from Bacillus sp. DSM 12649 disclosed as SEQ IDNO: 2 in WO 00/60060 (hereby incorporated by reference). Especiallypreferred are variants of the AA560 alpha-amylase, including the AA560variant disclosed in Example 7 and 8 (hereby incorporated by reference).

The amylases are normally incorporated in the detergent composition at alevel of from 0.000001% to 2% of enzyme protein by weight of thecomposition, preferably at a level of from 0.00001% to 1% of enzymeprotein by weight of the composition, more preferably at a level of from0.0001% to 0.5% of enzyme protein by weight of the composition, evenmore preferably at a level of from 0.001% to 0.2% of enzyme protein byweight of the composition.

Cellulases: Any cellulase suitable for use in alkaline solutions can beused. Suitable cellulases include those of bacterial or fungal origin.Chemically or genetically modified mutants are included. Suitablecellulases are disclosed in U.S. Pat. No. 4,435,307, which disclosesfungal cellulases produced from Humicola insolens. Especially suitablecellulases are the cellulases having colour care benefits. Examples ofsuch cellulases are cellulases described in European patent applicationNo. 0 495 257.

In a preferred embodiment the cellulase is a Thielavia terrestriscellulase, preferably the cellulase disclosed in SEQ ID NO: 9 in WO96/29397 and SEQ ID NO: 9 herein or an enzyme with at least 70% identitythereto. In a preferred embodiment cellulase is the Thielavia terrestrisvariant disclosed in Example 1 of WO 98/12307.

Commercially available cellulases include Celluzyme® produced by astrain of Humicola insolens, Carezyme® and Renozyme® (Novozymes A/S),and KAC-500(B)® (Kao Corporation). Cellulases are normally incorporatedin the detergent composition at a level of from 0.000001% to 2% ofenzyme protein by weight of the composition, preferably at a level offrom 0.00001% to 1% of enzyme protein by weight of the composition, morepreferably at a level of from 0.0001% to 0.5% of enzyme protein byweight of the composition, even more preferably at a level of from0.001% to 0.2% of enzyme protein by weight of the composition.

Mannanases: Any mannanase suitable for use in alkaline solutions can beused. Suitable mannanases include those of bacterial or fungal origin.Chemically or genetically modified mutants are included.

In a preferred embodiment the mannanase is derived from a strain of thegenus Bacillus, especially Bacillus sp. 1633 disclosed in positions31-330 of SEQ ID NO:2 or in SEQ ID NO: 5 of WO 99/64619 or Bacillusagaradhaerens, for example from the type strain DSM 8721. Further,suitable mannanases are Purabrite available from Genencor Inc. andMannawa® produced by Novozymes A/S.

Pectate Ivase: Any pectate lyase suitable for use in alkaline solutionscan be used. Suitable pectate lyases include those of bacterial orfungal origin. Chemically or genetically modified mutants are included.

In a preferred embodiment the pectate lyase is derived from a strain ofthe genus Bacillus, especially a strain of Bacillus subtilis, especiallyBacillus subtilis DSM14218 disclosed in SEQ ID NO:2 or a variant thereofdisclosed in Example 6 of WO 02/092741.

Peroxidases/Oxidases: Peroxidase enzymes are used in combination withhydrogen peroxide or a source thereof (e.g. a percarbonate, perborate orpersulfate). Oxidase enzymes are used in combination with oxygen. Bothtypes of enzymes are used for “solution bleaching”, i.e. to preventtransfer of a textile dye from a dyed fabric to another fabric when saidfabrics are washed together in a wash liquor, preferably together withan enhancing agent as described in e.g. WO 94/12621 and WO 95/01426.Suitable peroxidases/oxidases include those of plant, bacterial orfungal origin. Chemically or genetically modified mutants are included.

Peroxidase and/or oxidase enzymes are normally incorporated in thedetergent composition at a level of from 0.000001% to 2% of enzymeprotein by weight of the composition, preferably at a level of from0.00001% to 1% of enzyme protein by weight of the composition, morepreferably at a level of from 0.0001% to 0.5% of enzyme protein byweight of the composition, even more preferably at a level of from0.001% to 0.2% of enzyme protein by weight of the composition.

Mixtures of the above mentioned enzymes are encompassed herein, inparticular a mixture of two, three, four, five, six or more differentenzymes, for example a protease, an amylase, a lipase and a cellulase.

The enzyme of the invention, or any other enzyme incorporated in thedetergent composition, is normally incorporated in the detergentcomposition at a level from 0.000001% to 2% of enzyme protein by weightof the composition, preferably at a level from 0.00001% to 1% of enzymeprotein by weight of the composition, more preferably at a level from0.0001% to 0.5% of enzyme protein by weight of the composition, evenmore preferably at a level from 0.001% to 0.2% of enzyme protein byweight of the composition.

Enzymes may be incorporated in the form of liquid solutions orgranulates. Nondusting granulates may be produced, e.g., as disclosed inU.S. Pat. Nos. 4,106,991 and 4,661,452 (both to Novo Industri A/S, nowNovozymes A/S) and may optionally be coated by methods known in the art.Examples of waxy coating materials are poly(ethylene oxide) products(polyethyleneglycol, PEG) with mean molecular weights of 1000 to 20000;ethoxylated nonylphenols having from 16 to 50 ethylene oxide units;ethoxylated fatty alcohols in which the alcohol contains from 12 to 20carbon atoms and in which there are 15 to 80 ethylene oxide units; fattyalcohols; fatty acids; and mono- and di- and triglycerides of fattyacids. Examples of film-forming coating materials suitable forapplication by fluid bed techniques are given in GB 1483591.

Detergent composition examples: The detergent composition according tothe invention can be in liquid, paste, gels, bars, tablet or granularforms.

Granular compositions according to the present invention can also be in“compact form”, i.e. they may have a relatively higher density thanconventional granular detergents, i.e. form 550 to 950 g/l; in suchcase, the granular detergent compositions according to the presentinvention will contain a lower amount of “Inorganic filler salt”,compared to conventional granular detergents; typical filler salts arealkaline earth metal salts of sulphates and chlorides, typically sodiumsulphate; “Compact” detergent typically comprise not more than 10%filler salt. The liquid compositions according to the present inventioncan also be in “concentrated form”, in such case, the liquid detergentcompositions according to the present invention will contain a loweramount of water, compared to conventional liquid detergents. Typically,the water content of the concentrated liquid detergent is less than 30%,more preferably less than 20%, most preferably less than 10% by weightof the detergent compositions.

The compositions of the invention may for example, be formulated as handand machine laundry detergent compositions including laundry additivecompositions and compositions suitable for use in the pretreatment ofstained fabrics, rinse added fabric softener compositions, andcompositions for use in general household hard surface cleaningoperations and machine or hand dishwashing operations.

The following examples are meant to exemplify compositions for thepresent invention, but are not necessarily meant to limit or otherwisedefine the scope of the invention.

In the detergent compositions, the abbreviated component identificationshave the following meanings:

-   LAS: Sodium linear C₁₋₂ alkyl benzene sulphonate-   TAS: Sodium tallow alkyl sulphate-   XYAS: Sodium C_(1X)-C_(1Y) alkyl sulfate-   SS: Secondary soap surfactant of formula 2-butyl octanoic acid-   25EY: A C₁₂-C₁₅ predominantly linear primary alcohol condensed with    an average of Y moles of ethylene oxide-   45EY: A C₁₄-C₁₅ predominantly linear primary alcohol condensed with    an average of Y moles of ethylene oxide-   XYEZS: C_(1X)-C_(1Y) sodium alkyl sulfate condensed with an average    of Z moles of ethylene oxide per mole-   Nonionic: C₁₃-C₁₅ mixed ethoxylated/propoxylated fatty alcohol with    an average degree of ethoxylation of 3.8 and an average degree of    propoxylation of 4.5 sold under the tradename Plurafax LF404 by BASF    Gmbh-   CFM: C₁₂-C₁₄ alkyl N-methyl glucamide-   TFAA: C₁₈-C₁₈ alkyl N-methyl glucamide-   Silicate: Amorphous Sodium Silicate (SiO₂:Na₂O ratio=2.0)-   NaSKS-6: Crystalline layered silicate of formula δ-Na₂Si₂O₅-   Carbonate: Anhydrous sodium carbonate-   Phosphate: Sodium tripolyphosphate-   MA/M: Copolymer of 1:4 maleic/acrylic acid, average molecular weight    about 80,000-   Polyacrylate: Polyacrylate homopolymer with an average molecular    weight of 8,000 sold under the tradename Sokalan PA30 by BASF Gmbh-   Zeolite A: Hydrated Sodium Aluminosilicate of formula    Na₁₂(AlO₂SiO₂)₁₂.27H₂O having a primary particle size in the range    from 1 to 10 micrometers-   Citrate: Tri-sodium citrate dihydrate-   Citric: Citric Acid-   Perborate: Anhydrous sodium perborate monohydrate bleach, empirical    formula NaBO₂.H₂O₂-   PB4: Anhydrous sodium perborate tetrahydrate-   Percarbonate: Anhydrous sodium percarbonate bleach of empirical    formula 2Na₂CO₃.3H₂O₂-   TAED: Tetraacetyl ethylene diamine-   CMC: Sodium carboxymethyl cellulose-   DETPMP: Diethylene triamine penta (methylene phosphonic acid),    marketed by Monsanto under the Tradename Dequest 2060-   PVP: Polyvinylpyrrolidone polymer-   EDDS: Ethylenediamine-N,N′-disuccinic acid, [S,S] isomer in the form    of the sodium salt-   Suds 25% paraffin wax Mpt 50° C., 17% hydrophobic silica, 58%-   Suppressor: paraffin oil-   Granular Suds 12% Silicone/silica, 18% stearyl alcohol, 70%-   suppressor: starch in granular form-   Sulphate: Anhydrous sodium sulphate-   HMWPEO: High molecular weight polyethylene oxide-   TAE 25: Tallow alcohol ethoxylate (25)

Detergent Example I

A granular fabric cleaning composition in accordance with the inventionmay be prepared as follows: Sodium linear C₁₂ alkyl 6.5 benzenesulfonate Sodium sulfate 15.0 Zeolite A 26.0 Sodium nitrilotriacetate5.0 Enzymes (incl. endoglucanase) 0.1 PVP 0.5 TAED 3.0 Boric acid 4.0Perborate 18.0 Phenol sulphonate 0.1 Minors Up to 100

Detergent Example II

A compact granular fabric cleaning composition (density 800 g/l) inaccord with the invention may be prepared as follows: 45AS 8.0 25E3S 2.025E5 3.0 25E3 3.0 TFAA 2.5 Zeolite A 17.0 NaSKS-6 12.0 Citric acid 3.0Carbonate 7.0 MA/AA 5.0 CMC 0.4 Enzyme (incl. endo-glucanase) 0.1 TAED6.0 Percarbonate 22.0 EDDS 0.3 Granular suds suppressor 3.5 water/minorsUp to 100%

Detergent Example III

Granular fabric cleaning compositions in accordance with the inventionwhich are especially useful in the laundering of coloured fabrics wereprepared as follows: LAS 10.7 — TAS 2.4 — TFAA — 4.0 45AS 3.1 10.0 45E74.0 — 25E3S — 3.0 68E11 1.8 — 25E5 — 8.0 Citrate 15.0 7.0 Carbonate — 10Citric acid 2.5 3.0 Zeolite A 32.1 25.0 Na-SKS-6 — 9.0 MA/AA 5.0 5.0DETPMP 0.2 0.8 Enzyme (incl. endo-glucanase) 0.10 0.05 Silicate 2.5 —Sulphate 5.2 3.0 PVP 0.5 — Poly (4-vinylpyridine)-N- — 0.2Oxide/copolymer of vinyl- imidazole and vinyl- pyrrolidone Perborate 1.0— Phenol sulfonate 0.2 — Water/Minors Up to 100%

Detergent Example IV

Granular fabric cleaning compositions in accordance with the inventionwhich provide “Softening through the wash” capability may be prepared asfollows: 45AS — 10.0 LAS 7.6 — 68AS 1.3 — 45E7 4.0 — 25E3 — 5.0Coco-alkyl-dimethyl hydroxy- 1.4 1.0 ethyl ammonium chloride Citrate 5.03.0 Na-SKS-6 — 11.0 Zeolite A 15.0 15.0 MA/AA 4.0 4.0 DETPMP 0.4 0.4Perborate 15.0 — Percarbonate — 15.0 TAED 5.0 5.0 Smectite clay 10.010.0 HMWPEO — 0.1 Enzyme (incl. endo-glucanase) 0.10 0.05 Silicate 3.05.0 Carbonate 10.0 10.0 Granular suds suppressor 1.0 4.0 CMC 0.2 0.1Water/Minors Up to 100%

Detergent Example V

Heavy duty liquid fabric cleaning compositions in accordance with theinvention may be prepared as follows: I II LAS acid form — 25.0 Citricacid 5.0 2.0 25AS acid form 8.0 — 25AE2S acid form 3.0 — 25AE7 8.0 —CFAA 5 — DETPMP 1.0 1.0 Fatty acid 8 — Oleic acid — 1.0 Ethanol 4.0 6.0Propanediol 2.0 6.0 Enzyme (incl. endo-glucanase) 0.10 0.05 Coco-alkyldimethyl — 3.0 hydroxy ethyl ammonium chloride Smectite clay — 5.0 PVP2.0 — Water/Minors Up to 100%

Use of detergents: The enzyme composition of the invention may be usefulin a detergent composition for household or industrial laundering oftextiles and garments, and in a process for machine wash treatment offabrics comprising treating the fabrics during one or more washing cycleof a machine washing process with a washing solution containing theenzyme or enzyme preparation of the invention.

The enzyme composition of the invention may also be useful in adetergent composition for household or industrial dish or cutlery orother hard surface washing, and in a process for treatment of dishes,cutlery etc. comprising a treatment with a washing solution containingthe enzyme or enzyme preparation of the invention.

Typically, the detergent composition used in the washing processcomprises conventional ingredients such as surfactants (anionic,nonionic, zwitterionic, amphoteric), builders, bleaches (perborates,percarbonates or hydrogen peroxide) and other ingredients, e.g. asdescribed in WO 97/01629 which is hereby incorporated by reference inits entirety.

The endo-glucanase of the invention provides advantages such as improvedstain removal and decreased soil redeposition. Certain stains, forexample certain food stains, contain beta-glucans which make completeremoval of the stain difficult to achieve. Also, the cellulosic fibresof the fabrics may possess, particularly in the “non-crystalline” andsurface regions, glucan polymers that are degraded by this enzyme.Hydrolysis of such glucans, either in the stain or on the fabric, duringthe washing process decreases the binding of soils onto the fabrics.

Household laundry processes are carried out under a range of conditions.Commonly, the washing time is from 5 to 60 minutes and the washingtemperature is in the range 10-60° C., most commonly from 20-40° C.Prolonged soaking is commonly used. The washing solution is normallyneutral or alkaline, most commonly with pH 5-11.5. Bleaches are commonlyused, particularly for laundry of white fabrics and in the washing ofhard surfaces. These bleaches are commonly the peroxide bleaches, suchas sodium perborate, sodium percarbonate or hydrogen peroxide.

Materials & Methods

Strains and Donor Organism

The Bacillus sp. DSM 12648 mentioned above comprises the endo-glucanaseencoding DNA sequence shown in SEQ ID NO:1.

B.subtilis PL2306: This strain is the B.subtilis DN1885 with disruptedapr and npr genes (Diderichsen, B., Wedsted, U., Hedegaard, L., Jensen,B. R., Sjoholm, C. (1990) Cloning of aldB, which encodes alpha-acetolactate decarboxylase, an exoenzyme from Bacillus brevis. J.Bacteriol., 172, 4315-4321) disrupted in the transcriptional unit of theknown Bacillus subtilis cellulase gene, resulting in cellulase negativecells. The disruption was performed essentially as described in Eds. A.L. Sonenshein, J. A. Hoch and Richard Losick (1993) Bacillus subtilisand other Gram-Positive Bacteria, American Society for microbiology, p.618.

Competent cells were prepared and transformed as described by Yasbin, R.E., Wilson, G. A. and Young, F. E. (1975) Transformation andtransfection in lysogenic strains of Bacillus subtilis: evidence forselective induction of prophage in competent cells. J. Bacteriol,121:296-304.

General Molecular Biology Methods

Unless otherwise stated all the DNA manipulations and transformationswere performed using standard methods of molecular biology (Sambrook etal. (1989) Molecular cloning: A laboratory manual, Cold Spring Harborlab., Cold Spring Harbor, N.Y.; Ausubel, F. M. et al. (eds.) “Currentprotocols in Molecular Biology”. John Wiley and Sons, 1995; Harwood, C.R., and Cutting, S. M. (eds.) “Molecular Biological Methods forBacillus”. John Wiley and Sons, 1990).

Enzymes for DNA manipulations were used according to the manufacturer'sinstructions (e.g. restriction endonucleases, ligases etc. areobtainable from New England Biolabs, Inc.).

Plasmids

pMOL944. This plasmid is a pUB1 10 derivative essentially containingelements making the plasmid propagate in Bacillus subtilis, kanamycinresistance gene and having a strong promoter and signal peptide clonedfrom the amyL gene of B. licheniformis ATCC14580. The signal peptidecontains a SacII site making it convenient to clone the DNA encoding themature part of a protein in-fusion with the signal peptide. This resultsin the expression of a Preprotein which is directed towards the exteriorof the cell.

The plasmid was constructed by means of ordinary genetic engineering andis briefly described in the following.

Construction of PMOL944:

The pUB110 plasmid (McKenzie, T. et al., 1986, Plasmid 15:93-103) wasdigested with the unique restriction enzyme Ncil. A PCR fragmentamplified from the amyL promoter encoded on the plasmid pDN1981 (P. L.Jorgensen et al., 1990, Gene, 96, p3741.) was digested with Ncil andinserted in the Ncil digested pUB110 to give the plasmid pSJ2624.

The two PCR primers used have the following sequences: #LWN54945′-GTCGCCGGGGCGGCCGCTATCAATTGGTAACTGT (SEQ ID NO:5) ATCTCAGC-3′ #LWN54955′-GTCGCCCGGGAGCTCTGATCAGGTACCAAGCTTG (SEQ ID NO:6)TCGACCTGCAGAATGAGGCAGCAAGAAGAT-3′

The primer #LWN5494 inserts a NotI site in the plasmid.

The plasmid pSJ2624 was then digested with SacI and NotI and a new PCRfragment amplified on amyL promoter encoded on the pDN1981 was digestedwith SacI and NotI and this DNA fragment was inserted in the SacI-NotIdigested pSJ2624 to give the plasmid pSJ2670.

This cloning replaces the first amyL promoter cloning with the samepromoter but in the opposite direction. The two primers used for PCRamplification have the following sequences: #LWN59385′-GTCGGCGGCCGCTGATCACGTACCAAGCTTGTCG (SEQ ID NO:7)ACCTGCAGAATGAGGCAGCAAGAAGAT-3′ #LWN5939 5′-GTCGGAGCTCTATCAATTGGTAACTGTAT(SEQ ID NO:8) CTCAGC-3′

The plasmid pSJ2670 was digested with the restriction enzymes PstI andBcll and a PCR fragment amplified from a cloned DNA sequence encodingthe alkaline amylase SP722 (Patent # WO9526397-A1) was digested withPstI and Bcll and inserted to give the plasmid pMOL944. The two primersused for PCR amplification have the following sequence: #LWN78645′-AACAGCTGATCACGACTGATCTTTTAGCTT (SEQ ID NO:9) GGCAC-3′ #LWN79015′-AACTGCAGCCGCGGCACATCATAATGGGACAA (SEQ ID NO:10) ATGGG-3′

The primer #LWN7901 inserts a SacII site in the plasmid.

Genomic DNA Preparation

The strain DSM 12648 was propagated in liquid medium 2×TY containing 1%carboxymethyl-cellulose+(0.1M Na2CO3+0.1M NaHCO3 separately autoclavedand added aseptically after cooling to room temperature). After 16 hoursof incubation at 30° C. and 300 rpm, the cells were harvested, andgenomic DNA was isolated by the method described by Pitcher et al.[Pitcher, D. G., Saunders, N. A., Owen, R. J; Rapid extraction ofbacterial genomic DNA with guanidium thiocyanate; Lett Appl Microbiol1989, 8:151-156].

Media

TY (as described in Ausubel, F. M. et al. (eds.): “Current protocols inMolecular Biology”, John Wiley and Sons, 1995).

2xTY (as described in Ausubel, F. M. et al. (eds.): “Current protocolsin Molecular Biology”, John Wiley and Sons, 1995).

LB agar (as described in Ausubel, F. M. et al. (eds.): “Currentprotocols in Molecular Biology”, John Wiley and Sons, 1995).

LBPG is LB agar supplemented with 0.5% Glucose and 0.05 M potassiumphosphate, pH 7.0 AZCL-HE-cellulose is added to LBPG-agar to 0.5%AZCL-HE-cellulose is from Megazyme, Australia.

BPX media Is described in EP 0 506 780 (WO 91/09129).

Cal 18-2 media is described in patent application WO 00/75344 A1).

Test for Endo-Glucanase Activity

The purpose of this test, used in combination with the “test foranti-redeposition effect” described below, is to determine if an enzymeis an endo-glucanase with antiredeposition effect, according to thisinvention.

The test is made by determining the color released from an insoluble,colored, glucan substrate over a reaction time of 30 minutes at atemperature of 40° C.

Enzyme sample: The enzyme sample for testing is a solution of the enzymeprotein with a concentration of 0.1 mg/ml. Standard biochemicaltechniques can be used to verify the purity and determine the proteinconcentration of the sample.

Buffer: Prepare a 0.05M phosphate buffer solution with pH 7.0 fromNaH₂PO₄.2H₂O and NaOH and add 1 g/l nonionic surfactant (e.g. Berol 537,from Akzo Nobel).

Test method: Transfer 6 ml buffer into two test tubes. To one tube add50 μl of the enzyme sample. No enzyme is added to the second tube. Addone beta-glucazyme tablet (supplied by Megazyme, Ireland, cataloguenumber T-BGZ) to each tube. Mix the contents of the tubes for 10 secondswith a vortex mixer, then place the tubes in a 40° C. water bath. After10 minutes and after 20 minutes, mix the contents of the tubes byinverting the tubes. After 30 minutes, mix the contents of the tubes byinverting the tubes and then filter the contents of the tubes through aWhatman GF/C 9 cm filter, collecting the filtrates in clean tubes.Measure the color released as OD at 590 nm using a spectrophotometer.Calculate deltaOD by subtracting the result with no enzyme from theresult with enzyme.

If deltaOD>0.2 then the enzyme is an endo-glucanase.

Test for Anti-Redeposition Effect

The purpose of this test, used in combination with the “test forendo-glucanase activity” described above, is to determine if an enzymeis an endo-glucanase with antiredeposition effect, according to thisinvention.

The anti-redeposition effect is measured by a wash test. Theanti-redeposition wash test is made by washing samples of soiled cottonfabric and samples of clean cotton fabric, both together, in asmall-scale wash test apparatus. After the washing the soil on theoriginally clean cotton fabric is evaluated by light reflectance.

Enzyme sample: The enzyme sample for testing is a solution of the enzymeprotein with a concentration of 0.1 mg/ml. Standard biochemicaltechniques can be used to verify the purity and determine the proteinconcentration of the sample.

Cotton fabric: #2003 white woven 100% cotton fabric, supplied byTanigashira, 4-11Komatsu Yodogawa-ku, Osaka, 533-0004, Japan. The newcotton fabric is pre-washed three times before use in the wash test. Thepre-washing is done using a European household front-loader washingmachine, and using a standard 40° C. wash process. LAS (Surfac® SDBS80sodium alkylbenzene sulfonate, 80%) is added to the wash water atconcentration 0.5 g per liter and the wash solution pH is adjusted to 10by addition of sodium carbonate. After the pre-washing the fabric isdried in a tumbler drier. Swatches of the pre-washed cotton fabric, size5×5 cm, weight approximately 0.3 g each, are cut out and these swatchesare used for the wash tests.

Soiled cotton swatches: Swatches of the pre-washed #2003 fabric,prepared as above, are soiled as follows. A suspension of 210 mg carbonblack (“carbon for detergency tests”, supplied by Sentakukagaku-kyokai,2-11-1 Shimomaruko Ohta-ku, Tokyo 146-8620, Japan) is prepared in 75 mltetrachloroethylene (Fluka, cat. nr. 86972) by strong stirring. Thecotton swatches are placed flat on a horizontal metal surface. 300 μl ofthe carbon suspension is pipetted onto the centre of each cotton swatch.The soiled cotton swatches are allowed to dry at room temperatureovernight.

Detergent solutions: Detergent solutions are prepared as follows. Toprepare 1 liter of solution, dissolve in deionised water 0.5 g sodiumcarbonate and 1.0 g sodium hydrogen carbonate and add 2 ml of a solutioncontaining 117.8 g/l CaCl₂.2H₂O and 54.3 g/l MgCl₂.6H₂O. Thiscalcium/magnesium addition provides a water hardness of 12° dH. Add 0.5g LAS (Surfac SDBS80 sodium alkylbenzene sulfonate, 80%, supplied bySurfachem, UK or an equivalent material) and adjust the final volume to1 liter. Adjust the pH to 9.5 (e.g. by addition of sodium carbonate)Wash tests: Three soiled swatches (prepared as described) and threeclean swatches (of the same pre-washed #2003 cotton) are washed in aMini-Terg-O-Tometer machine. The Mini-Terg-O-Tometer is a small-scaleversion of the Terg-O-Tometer test washing machine described in Jay C.Harris, “Detergency Evaluation and Testing”, Interscience PublishersLtd. (1954) pp. 60-61. The following conditions are used: Beaker size250 ml Wash solution volume 100 ml Wash temperature 40° C. Wash time 30minutes Agitation 150 rpm

The detergent solutions are pre-warmed to 40° C. before starting thetest. The cotton swatches and 100 μl of the enzyme sample are added atthe start of the 30 minute wash period. The reference test, with noadded enzyme, is started at the same time.

After the wash, the fabric swatches are rinsed for 5 minutes underrunning tap water, then spread out flat and allowed to air dry at roomtemperature overnight.

Instrumental evaluations: Light reflectance evaluation of the originallyclean fabric swatches is done using a Macbeth Color Eye 7000 reflectancespectrophotometer. The measurements are made at 500 nm. The UV filter isnot included. Measurements are made on the front and back of eachswatch. An average result for reflectance (R, 500 nm) for the threeoriginally clean swatches is then calculated from the six measurements.

The anti-redeposition effect of a given enzyme sample is calculated as:

(R, 500 nm with the enzyme)-(R, 500 nm no enzyme added)

The enzyme is an endo-glucanase with anti-redeposition effect accordingto the invention if the anti-redeposition effect obtained is >7.5, andthe result in the test for endoglucanase activity, above, is >0.2.

LAS Stability Test

The test is made by comparing the color released from an insolublecolored cellulase substrate over a reaction time of 60 minutes at atemperature of 40° C. when the solution contains a buffer but nosurfactant and when the solution contains a buffer and LAS. If theenzyme is stable during the 60 minute reaction time then the colorreleased is greater than if the enzyme is not stable, because the stableenzyme will continue to degrade the substrate whereas the non-stableenzyme will not continue to degrade the substrate. In the context of thepresent invention, if the result with LAS is >50% of the result withoutLAS then the enzyme is LAS stable.

The details of the test method are as follows.

Buffer (no LAS):

-   Prepare a 0.05M phosphate buffer solution with pH 7.0 from    NaH₂PO₄.2H₂O and NaOH.-   Buffer (with LAS):-   Dissolve 1.0 g LAS in 1 liter of the above buffer. The LAS is    Surface SDBS80 sodium alkylbenzene sulfonate, 80% (supplied by    Surfachem, UK) or an equivalent material.    Test Method:-   Prepare a dilution of the enzyme to be tested in deionised water.    The concentration of the enzyme is such that the result for deltaOD    (no LAS), calculated as described below, is in the range 0.2 to 0.5.

Transfer 6 ml of the buffer (no LAS) into two test tubes. Add 150 μl ofthe enzyme dilution into one tube. No enzyme is added to the other tube.Add one Cellazyme® C tablet (supplied by Megazyme, Ireland, cataloguenumber T-CCZ) into both tubes. Mix strongly for about 10 seconds using avortex mixer. Place both tubes in a 40° C. water bath. After 15, 30 and45 minutes, mix the contents of the tubes by inverting the tubes. After60 minutes, mix the contents of the tubes by inverting the tubes andthen filter the contents through a 9 cm diameter Whatman® GF/C filter,collecting the filtrate in a clean tube. Measure the OD of the filtrateat 590 nm using a spectrophotometer. Calculate deltaOD (no LAS) bysubtracting the result with no enzyme from the result with enzyme.

Then repeat the test using the same concentration of the enzyme and with6 ml buffer (with LAS) instead of 6 ml buffer (no LAS). CalculatedeltaOD (with LAS) by subtracting the result with no enzyme from theresult with enzyme.

Calculate the LAS stability %:(deltaOD (with LAS)/deltaOD(no LAS))*100Wash Tests with the Endo-Glucanase In Combination with Other Enzymes

This procedure is used to determine the enzyme detergency benefitvalues, see Examples 6-11 below.

The wash tests are made by washing samples of soiled cotton fabric andsamples of clean cotton fabric, both together, in a small-scale washtest apparatus. After the washing the soil on the cotton fabric isevaluated by light reflectance. Both the originally soiled cotton fabricand the originally clean cotton fabric samples are evaluated.

Cotton fabric: #2003 white woven 100% cotton fabric, supplied byTanigashira, 4-11Komatsu Yodogawa-ku, Osaka, 533-0004, Japan. The newcotton fabric is pre-washed three times before use in the wash test. Thepre-washing is done using a European household front-loader washingmachine, and using a standard 40° C. wash process. LAS (Surfac® SDBS80sodium alkylbenzene sulfonate, 80%) is added to the wash water atconcentration 0.5 g per liter and the wash solution pH is adjusted to 10by addition of sodium carbonate. After the pre-washing the fabric isdried in a tumbler drier. Swatches of the pre-washed cotton fabric, size5×5 cm, weight approximately 0.3 g each, are cut out and these swatchesare used for the wash tests.

Soiled cotton swatches: These are prepared from the 5×5 cm swatchesdescribed above.

Wash tests: Three soiled swatches and three clean swatches are washed ina MiniTerg-O-Tometer machine. The Mini-Terg-O-Tometer is a small-scaleversion of the Terg-OTometer test washing machine described in Jay C.Harris, “Detergency Evaluation and Testing”, Interscience PublishersLtd. (1954) pp. 60-61. The following conditions are used: Beaker size250 ml Wash solution volume 100 ml Wash temperature 40° C. Wash time 30minutes Agitation 150 rpm

The detergent solutions are pre-warmed to 40° C. before starting thetest. The fabric and the enzymes are added at the start of the 30 minutewash period. After the wash, the fabric swatches are rinsed for 5minutes under running tap water, then spread out flat and allowed to airdry at room temperature overnight.

Instrumental evaluations: Light reflectance evaluation of the fabricswatches is done using a Macbeth Color Eye 7000 reflectancespectrophotometer. The measurements are made at 500 nm. The UV filter isnot included. Measurements are made on the front and back of eachswatch. The soiled swatches are measured in the centre of the soiledarea. Average results for reflectance (R, 500 nm) for the soiledswatches and for the clean swatches are then calculated from the sixmeasurements on each type.

Detergent solutions: Detergent solutions are prepared as follows: Toprepare 1 liter of solution, dissolve in deionised water 0.5 g sodiumcarbonate and 11.0 g sodium hydrogen carbonate and add 2 ml of asolution containing 117.8 g/l CaCl₂.2H₂O and 54.3 g/l MgCl₂.6H₂O. Thiscalcium/magnesium addition provides a water hardness of 12° dH. Addnonionic surfactant (Berol® 537, Akzo Nobel) and/or LAS (Surfac® SDBS80sodium alkylbenzene sulfonate, 80%) and adjust the final volume to 1liter. Adjust the pH to either pH 9.5 (by addition of sodium carbonate)or to pH 7.5 (by addition of 10% citric acid solution).

The concentration of LAS and of nonionic surfactant and the detergentsolution pH are specified below for each wash test.

Enzyme addition: The enzymes to be tested are pre-dissolved at knownconcentrations in water, and the required amount of enzyme is added tothe detergent solution at the start of the wash process.

Calculation of enzyme detergency benefit: The enzyme detergency benefitis a measure of how much more clean the swatches, both the originallysoiled and the originally clean, become as a result of including enzymesin the wash test. The enzyme detergency benefit is calculated asfollows: After the wash test the average R, 500 nm value for the soiledswatches is R, soiled. After the wash test the average R, 500 nm valuefor the clean swatches is R, clean. The enzyme detergency benefit from awash test with enzymes is the sum of R, soiled+R1 clean with enzymesminus the sum of R, soiled+R, clean with no added enzyme.

The enzyme detergency benefit value determined in this way is a combinedmeasure both of the removal of soil from the fabric and of theredeposition of soil onto the fabric. Thus the enzyme detergency benefitvalue can have values that are negative or positive. The enzymedetergency benefit value can be used to compare the performance ofdifferent enzymes. The highest positive detergency benefit value is thepreferred result.

EXAMPLE 1

Cloning and Expression of Endo-Glucanase Gene from Bacillus sp.

Sub-Cloning and Expression of Mature Endo-Glucanase in B. subtilis.

The endo-glucanase encoding DNA sequence of the invention was PCRamplified using the PCR primer set consisting of these twooligo-nucleotides: #168684 5′-CAT TCT GCA GCC GCG GCA GCA GAA (SEQ IDNO:11) GGA AAC ACT CGT GAA GAC-3′ #168685 5′-GCG TTG AGA CGC GCG GCC GCTTAC (SEQ ID NO:12) TCT TCT TTC TCT TCT TTC TC-3′

Restriction sites SacI and NotI are underlined.

The oligonucleotides were used in a PCR reaction in HiFidelity™ PCRbuffer (Boehringer Mannheim, Germany) supplemented with 200 micro M ofeach dNTP, 2.6 units of HiFidelity™ Expand enzyme mix and 200 pmol ofeach primer. Chromosomal DNA isolated from Bacillus sp. DSM12648 asdescribed above was used as template.

The PCR reaction was performed using a DNA thermal cycler (Landgraf®,Germany). One incubation at 94° C. for 1 min followed by ten cycles ofPCR performed using a cycle profile of denaturation at 94° C. for 15sec, annealing at 60° C. for 60 sec, and extension at 72° C. for 120sec, followed by twenty cycles of denaturation at 94° C. for 15 sec, 60°C. for 60 sec and 72° C. for 120 sec (at this elongation step 20 sec areadded every cycle). 5 μl aliquots of the amplification product wasanalysed by electrophoresis in 0.7% agarose gels (NuSieve®, FMC). Theappearance of a DNA fragment size 2.4 kb indicated proper amplificationof the gene segment.

Subcloning of PCR Fragment:

45 microL aliquots of the PCR products generated as described above werepurified using QiAquick® PCR purification kit (Qiagen®, USA) accordingto the manufacturer's instructions. The purified DNA was eluted in 50microL of 10 mM Tris-HCl, pH 8.5. 5 μg of pMOL944 and 25 microL of thepurified PCR fragment was digested with SacII and NotI, electrophoresedin 0.7% agarose gels (NuSieve®, FMC), the relevant fragments wereexcised from the gels, and purified using QlAquick® Gel extraction Kit(Qiagen®, USA) according to the manufacturers instructions. The isolatedPCR DNA fragment was then ligated to the SacII-NotI digested andpurified pMOL944. The ligation was performed overnight at 16° C. using0.5 micro g of each DNA fragment, 1 U of T4 DNA ligase and T4 ligasebuffer (Boehringer Mannheim, Germany).

The ligation mixture was used to transform competent B. subtilis PL2306.The transformed cells were plated onto LBPG-10 micro g/ml ofkanamycin-agar plates. After 18 hours incubation at 37° C. colonies wereseen on the plates. Several clones were analyzed by isolating plasmidDNA from overnight culture broths.

One such positive clone was re-streaked several times on agar plates asused above; this clone was called MB1181-7. The clone MB1181-7 was grownovernight in TY-10 micro g/mL kanamycin at 37° C., and next day 1 ml ofcells were used to isolate a plasmid from the cells using the Qiaprep®Spin Plasmid Miniprep Kit #27106 according to the manufacturersrecommendations for B. subtilis plasmid preparations. This DNA wassequenced and revealed a DNA sequence identical to the endo-glucanasegene in SEQ ID NO:1 bp 1-2322 encoding the mature endo-glucanase. Thederived protein sequence is represented in SEQ ID NO: 2.

EXAMPLE 2

Expression and Recovery of the Endo-Glucanase from Bacillus sp. DSM12648

MB1181-7 obtained as described in Example 1 was grown in 15×200 mlCal-18-2 media with 10 microg/mL of kanamycin, in 500 mL two-baffledshake flasks, for 4 days at 37° C. at 300 rpm, whereby about 2500 ml ofculture broth was obtained. The culture fluid was flocculated by adding50% CaCl₂ (10 ml per liter of culture broth) together with 11% sodiumaluminate (10 mL per liter of culture broth), maintaining the pH between7.0 and 7.5 by adding 20% formic acid. Cationic agent Superfloc® C521(25 mL of a 10% v/v dilution per liter of culture broth) and anionicagent Superfloc® A130 (75 ml of a 0.1% w/v dilution in water per literof culture broth) was added during agitation to complete theflocculation. The flocculated material was separated by centrifugationusing a Sorval® RC 3B centrifuge at 10000 rpm for 30 min at 6° C. Theresulting supernatant contained the endo-glucanase activity.

The supernatant was clarified using Whatman glass filters GF/D and C.Then ultrafiltration was used to concentrate and reduce the ionicstrength of the solution. The ultrafiltration membrane was Filtron® UFwith a cut-off of 10 kDa. After ultra-filtration the solution hadconductivity<3 mS/cm. The pH was adjusted to pH 8.0.

Anion-exchange chromatography on Q-Sepharose® was then used foradditional purification. The solution from ultra-filtration was appliedto a 300 mL column containing QSepharose® (Pharmacia) equilibrated witha buffer of 25 mmol Tris pH 8.0. The endoglucanase bound to theQ-Sepharose®, and was then eluted using a 0.5 M NaCl gradient. Thefractions with high endo-glucanase activity were pooled. Theendo-glucanase activity of the final pooled endo-glucanase solution wasapproximately 1000 ECU per mL.

The activity units, ECU, are determined by method SM-0302, which isavailable from Novozymes A/S on request. In the ECU method the abilityof the enzyme sample to reduce the viscosity of a solution ofcarboxymethyl-cellulose is determined, and the result is given in ECU.Conditions: CMC type 7LFD from Hercules, pH7.5 in 0.1M phosphate buffer,CMC concentration 31.19 per liter, reaction at 40° C. for 30 minutes. Avibration viscosimeter such as MIVI 3000, Sofraser®, France is used tomeasure the viscosity.

EXAMPLE 3 Stain Removal and Anti-Redeposition Effect

This test demonstrates the stain removal and anti-redeposition effectsof the endoglucanase obtained in Example 2. Additionally this testdemonstrates that the enzyme performance is essentially unchanged whensodium perborate bleach is included.

Cotton swatches are stained with beta-glucan (from barley) plus carbonblack. Soiled swatches are washed together with clean swatches. Afterwashing the swatches are rinsed and dried. The soil removal from thesoiled switches and the soil redeposition onto the clean swatches isdetermined by reflectance measurements. The soil removal and soilredeposition after washing without or with addition of theendo-glucanase are compared.

Swatches: Cut from 100% cotton fabric, type #2003 (Tanigashira, Osaka,Japan), pre-washed at 40° C. as a precaution to remove any water solublecontaminations, size 5×5 cm, weight approximately 0.3 g.

Washing equipment: Stirred beakers, beaker volume 250 ml, withtemperature control by water bath heating. The equipment is theMini-Terg-O-Tometer, a multi-beaker miniature agitator washer, asdescribed in the Materials and Methods section.

Detergent solution: Prepared by adding the following into deionisedwater.

Sodium carbonate, 0.5 g per liter

Sodium bicarbonate, 0.7 g per liter

Ca²⁺/Mg²⁺, to give water hardness 12“dH

Anionic surfactant, Surface SDBS80 (sodium alkylbenzene sulphonate), 0.5g per liter

Nonionic surfactant, Berol® 537 (Akzo Nobel), 1.0 g per liter

Sodium perborate, type SPB from wfk Testgewebe, either 0 or 1.0 g perliter

Solution pH is approximately 9.5.

Washing procedure: 100 mL detergent solution is added to each beaker.The water bath temperature is 40° C. The mechanical agitators areoperated at approximately 125 rpm. The detergent solutions arepre-warmed for 10 minutes and then the endo-glucanase and the swatchesare added. In each case three soiled swatches (prepared as describedbelow) and three clean swatches are added to each beaker. After washingfor 30 minutes, the swatches are removed from the detergent solution,rinsed under running tap water for 5 minutes, spread flat on absorbentpaper and allowed to dry.

Reflectance measurements: Made using a Macbeth® 7000 Color Eyereflectance spectrophotometer. In the case of the soiled swatches, eachswatch is measured once in the center of the soiled area, then theaverage value is calculated. In the case of the clean swatches, eachswatch is measured once on each side, then the average value iscalculated. The reflectance measurements are all made at 500 nm.

Soiled swatches: Soiled swatches are made using beta-glucan (frombarley) and carbon black (“carbon for detergency tests” supplied bySentaku Kagaku Kyokai, Tokyo, Japan). Dissolve about 0.67 g ofbeta-glucan in 100 mL tap water by stirring and warming to >50° C. Add0.33 g carbon black. Blend with an UltraTurraxe T25 blender, speed 4000rpm for 2 minutes. Apply 250 microL of the beta-glucan/carbon onto thecenter of each swatch. Allow to dry overnight at room temperature.

The swatches used in this example had an average reflectance value of93.5 before soil application and 17.5 after soiling.

Endo-glucanase addition: The endo-glucanase from Example 2 was added togive an activity concentration of 0, 20 or 100 ECU per liter ofdetergent solution. The activity units, ECU, are determined by methodSM-0302, as above.

Results: Detergent without bleach (average of reflectance measurementsafter washing) Endo-glucanase added Soiled swatches Clean swatches 025.1 33.5  20 ECU per liter 35.7 46.7 100 ECU per liter 40.2 59.1

Results: Detergent with bleach (average of reflectance measurementsafter washing) Endo-glucanase added Soiled swatches Clean swatches 024.6 27.7  20 ECU per liter 36.8 52.6 100 ECU per liter 39.3 63.2

The endo-glucanase increased the removal of soil from the fabric, asseen by the increased reflectance value of the stained swatches afterwashing with endo-glucanase as compared to the result after washingwithout endo-glucanase. The endo-glucanase also decreases the soilredeposition, as seen by the increased reflectance value of the cleanswatches after washing with endo-glucanase. The improvements of soilremoval and antiredeposition provided by the endo-glucanase areessentially unchanged by the addition of the bleach.

EXAMPLE 4 Anti-Redeposition Effect

Clean cotton fabric is washed together with soiled cotton fabric in asolution of a household detergent. The wash is carried out in aTerg-O-Tometer. During the wash, soil is released from the soiled fabricinto the detergent liquor. This soil can then redeposit onto the cleancotton. After washing, the cotton fabrics are rinsed and dried, and thenmeasured with a reflectance spectrophotometer in order to detect thedegree of soil redeposition.

-   Detergent: Powder household detergent, Asian.-   Detergent concentration: 0.67 g/l in water with hardness 4° d H.-   1000 mL of detergent solution per T-O-T beaker.-   Cotton fabric: Total of 33 g fabric per T-O-T beaker, comprising    suitably sized pieces of:    -   white woven cotton, #2003 (Tanigashira, Osaka, Japan), total        weight 11 g    -   white cotton interlock, total weight 13 g    -   soiled cotton fabric, type EMPA101 (EMPA, Switzerland), total        weight 9 g.-   Wash: Temperature 25° C., wash time 40 minutes, at 125 rpm. After    washing the #2003 cotton is rinsed under running tap water for 10    minutes, then dried.

Reflectance measurements. The pieces of #2003 woven cotton are measured,on both sides, using a Macbeth® 7000 reflectance spectrophotometer, 500nm. The average result for measurements from each T-O-M beaker iscalculated.

Enzyme addition: In this trial, the glucanase prepared as described inExample 2 was added to the detergent liquor before the start of the washstep.

Results: Endo-glucanase added Reflectance of #2003, (ECU per liter) at500 nm 0 76.67 0 76.05 1 81.86 5 84.30 20 84.85 50 85.99

The activity units, ECU, are determined by method SM-0302, as above.From the results it can be concluded that addition of the endo-glucanasereduces the soil redeposition.

Example 5 LAS Stability of Enzyme Samples

The LAS stability % was determined by the procedure described above. Thefollowing exzymes were tested:

Carezyme is the trade name for enzyme products from Novozymes A/S thatcontain (as the only enzyme component) the 43 kD cellulase derived fromHumicola insolens DSM 1800 disclosed in WO 91/17243 (SEQ ID NO: 2 andhereby incorporated by reference). Detergent compositions containingthis enzyme are disclosed in EP 822 973.

Renozyme is the trade name for enzyme products from Novozymes A/S thatcontain (as the only enzyme component) the Thielavia terrestriscellulase variant disclosed in Example 1 of WO 98/12307

Endo-glucanase. This is an endo-glucanase with anti-redeposition effectas defined above. A method for preparation of this enzyme is describedin Example 2. Results: Enzyme LAS stability % Carezyme 13 Renozyme 62Endo-glucanase 83

These results show that, according to the definition used, Carezyme isnot LAS stable and that both Renozyme and the endo-glucanase are LASstable.

EXAMPLE 6 Wash test: Renozyme® and Endoglucanase

The purpose of this test was to measure the enzyme detergency benefit ofRenozyme® alone and of Renozyme® in combination with endo-glucanase.

For this test the soiled swatches were prepared as follows: Swatches ofthe prewashed #2003 fabric, prepared as above, were soiled as follows. Asuspension of 210 mg carbon black (“carbon for detergency tests”,supplied by Sentakukagaku-kyokai, 2-11-1 Shimomaruko Ohta-ku, Tokyo146-8620, Japan) was prepared in 75 ml tetrachloroethylene (Fluka®, cat.nr. 86972) by strong stirring. The cotton swatches were placed flat on ahorizontal metal surface. 300 μl of the carbon suspension was pipettedonto the centre of each cotton swatch. The soiled cotton swatches wereallowed to dry at room temperature overnight.

For this test the detergent composition was as described above and withthe following surfactants and pH:

-   -   LAS: 0.5 g per liter    -   Nonionic: none    -   pH: 9.5

Enzyme detergency benefit results: Renozyme ® at Renozyme ® at 50 μg/l125 μg/l No endo-glucanase 6.1 8.1 endo-glucanase at 75 μg/l 15.4endo-glucanase at 190 μg/l 16.9

The enzyme concentrations are specified here in terms of the equivalentconcentration of the pure, catalytically active enzyme proteins, inorder to avoid ambiguity resulting from activity assay procedures.Standard biochemical techniques can be used to purify and characterisethe enzymes.

A suitable cellulase can be obtained from samples of the commercialproduct Renozyme” from Novozymes A/S or derived as described in Example1 of WO 98/12307. The endo-glucanase is an endo-glucanase withanti-redeposition effect as defined above. A method for preparation ofthis enzyme is described in Example 2.

The results show that the combinations of Renozyme® and theendo-glucanase give a higher enzyme detergency benefit than Renozyme®alone in a detergent solution that contains LAS but no othersurfactants.

EXAMPLE 7 Wash test: Stainzyme® and Endo-Glucanase

The purpose of this test was to measure the enzyme detergency benefit ofStainzyme® alone and of Stainzyme® in combination with endo-glucanase ona soil that contains starch.

Stainzyme® is the trade name of a commercially available alpha-amylaseproduct produced by Novozymes A/S. Stainzyme® is intended for use indetergents for laundry and hard surface cleaning.

For this test the soiled swatches were prepared as follows: Swatches ofthe prewashed #2003 fabric, prepared as above, were soiled as follows. Asuspension of 4.5 g of potato starch (“kartoffelmel”, produced by KMCkartoffelmel-centralen, DK-7400 Herning, Denmark, starch contentapproximately 80%) was prepared in 500 ml tap water with stirring, thenheated to boiling and then cooled. To 100 ml of this solution, 332 mg ofcarbon black (“carbon for detergency tests”, supplied bySentakukagaku-kyokai, 2-11-1 Shimomaruko Ohtaku, Tokyo 146-8620, Japan)were added, and the suspension was homogenised with an UltraTurrax®blender. The cotton swatches were placed flat on a horizontal metalsurface. 250 μl of the carbon/starch suspension was pipetted onto thecentre of each cotton swatch. The soiled cotton swatches were allowed todry at room temperature overnight.

For this test the detergent composition was as described above and withthe following surfactants and pH:

-   -   LAS: 0.4 g per liter    -   Nonionic: 0.2 g per liter    -   pH: 9.5

Enzyme detergency benefit results: Stainzyme ® Stainzyme ® at 2.1 mg/lat 5.3 mg/l No endo-glucanase −2.9 1.7 endo-glucanase at 75 μg/l 10.1endo-glucanase at 190 μg/l 13.1

The enzyme concentrations are specified here in terms of the equivalentconcentration of the pure, catalytically active enzyme proteins, inorder to avoid ambiguity resulting from activity assay procedures.Standard biochemical techniques can be used to purify and characterisethe enzymes.

A suitable amylase can be obtained from samples of the commercialproduct Stainzyme® from Novozymes A/S or derived from B. licheniformisvariants as disclosed in WO 01/66712 and WO 01/64852.

The endo-glucanase is an endo-glucanase with anti-redeposition effect asdefined above. A method for preparation of this enzyme is described inExample 2.

In this test the soil contains starch. The results show that thecombinations of endoglucanase and Stainzyme® give a higher enzymedetergency benefit than Stainzyme” alone.

EXAMPLE 8 Wash test: Savinase® and Endo-Glucanase

The purpose of this test was to measure the enzyme detergency benefit ofSavinase alone and of Savinase in combination with endo-glucanase on asoil that contains protein.

Savinase® is the trade name of subtilisin 309. It is a commerciallyavailable protease product produced by Novozymes A/S. Savinase® isintended for use in detergents for laundry and hard surface cleaning.

For this test the soiled swatches were prepared as follows: Swatches ofthe prewashed #2003 fabric, prepared as above, were soiled as follows. Asolution of 7.2 g of gelatine (gelatine from porcine skin, Fluka®, cat.number 04055) was prepared in 100 ml tap water by warming and withstirring. 10 ml of this solution was diluted to 50 ml with water and 165mg of carbon black (“carbon for detergency tests”, supplied bySentakukagaku-kyokai, 2-11-1 Shimomaruko Ohta-ku, Tokyo 146-8620, Japan)were added, and the suspension was homogenised with an UltraTurrax®blender. The cotton swatches were placed flat on a horizontal metalsurface. 250 μl of the carbon/gelatine suspension was pipetted onto thecentre of each cotton swatch. The soiled cotton swatches were allowed todry at room temperature overnight.

For this test the detergent composition was as described above and withthe following surfactants and pH:

-   -   LAS: 0.4 g per liter    -   Nonionic: 0.2 g per liter    -   pH: 9.5

Enzyme detergency benefit results: Savinase ® Savinase ® at 0.8 mg/l at1.9 mg/l No endo-glucanase 3.7 7.6 Endo-glucanase at 75 μg/l 16.6Endo-glucanase at 190 μg/l 17.7

The enzyme concentrations are specified here in terms of the equivalentconcentration of the pure, catalytically active enzyme proteins, inorder to avoid ambiguity resulting from activity assay procedures.Standard biochemical techniques can be used to purify and characterisethe enzymes.

A suitable protease can be obtained from samples of the commercialproduct Savinase® from Novozymes A/S or derived from Bacillus lentusNCIB 10309 as described in U.S. Pat. No. 3,723,250.

The endo-glucanase is an endo-glucanase with anti-redeposition effect asdefined above. A method for preparation of this enzyme is described inExample 2.

In this test the soil contains protein. The results show that thecombinations of the endo-glucanase and Savinase® give a higher enzymedetergency benefit than Savinase® alone.

EXAMPLE 9 Wash test: Mannanase and Endo-Glucanase

The purpose of this test was to measure the enzyme detergency benefit ofa mannanase alone and of a mannanase in combination with endo-glucanaseon a soil that contains galactomannan.

For this test the soiled swatches were prepared as follows: Swatches ofthe prewashed #2003 fabric, prepared as above, were soiled as follows. Asolution of 667 mg of guar gum (Sigma®, G 4129) was prepared in 200 mltap water by warming and with stirring and 332 mg of carbon black(“carbon for detergency tests”, supplied by Sentakukagaku-kyokai, 2-11-1Shimomaruko Ohta-ku, Tokyo 146-8620, Japan) were added, and thesuspension was homogenised with an UltraTurrax® blender. The cottonswatches were placed flat on a horizontal metal surface. 500 μl of thecarbon/guar gum suspension was pipetted onto the centre of each cottonswatch. The soiled cotton swatches were allowed to dry at roomtemperature overnight.

For this test the detergent composition was as described above and withthe following surfactants and pH:

-   -   LAS: 0.4 g per liter    -   Nonionic: 0.2 g per liter    -   pH: 9.5

Enzyme detergency benefit results: Mannanase at Mannanase at 0.3 mg/l0.75 mg/l No endo-glucanase 4.8 6.2 Endo-glucanase at 75 μg/l 16.2Endo-glucanase at 190 μg/l 12.6

The enzyme concentrations are specified here in terms of the equivalentconcentration of the pure, catalytically active enzyme proteins, inorder to avoid ambiguity resulting from activity assay procedures.Standard biochemical techniques can be used to purify and characterisethe enzymes.

A suitable mannanase can be obtained from Bacillus sp.1633, disclosed inWO 99/64619. Preferred is the mannanase of SEQ ID NO: 2 of WO 99/64619.

The endo-glucanase is an endo-glucanase with anti-redeposition effect asdefined above. A method for preparation of this enzyme is described inExample 2.

In this test the soil contains a galactomannan. The results show thatthe combinations of the endo-glucanase and mannanase give a higherenzyme detergency benefit than mannanase alone.

EXAMPLE 10 Wash test: Pectate Lyase and Endo-Glucanase

The purpose of this test was to measure the enzyme detergency benefit ofa pectate lyase (BioPrep® L) alone and in combination withendo-glucanase on a soil that contains pectin.

BioPrep® is the trade name of a commercially available pectate lyaseproduct produced by Novozymes A/S. BioPrep® is intended for use incleaning of cotton fibres.

For this test the soiled swatches were prepared as follows: Swatches ofthe prewashed #2003 fabric, prepared as above, were soiled as follows. Asolution of 150 m g of pectin (Sigma®, P 9311) was prepared in 50 ml tapwater by warming and with stirring and 83 mg of carbon black (“carbonfor detergency tests”, supplied by Sentakukagaku-kyokai, 2-11-1Shimomaruko Ohta-ku, Tokyo 146-8620, Japan) were added, and thesuspension was homogenised with an UltraTurrax® blender. The cottonswatches were placed flat on a horizontal metal surface. 500 μl of thecarbon/pectin suspension was pipetted onto the centre of each cottonswatch. The soiled cotton swatches were allowed to dry at roomtemperature overnight.

For this test the detergent composition was as described above and withthe following surfactants and pH:

-   -   LAS: 0.4 g per liter    -   Nonionic: 0.2 g per liter    -   pH: 9.5

Enzyme detergency benefit results: BioPrep ® at 1 mg/l BioPrep ® at 2.5mg/l No endo-glucanase −8.0 −5.1 Endo-glucanase at 75 μg/l 6.6Endo-glucanase at 190 μg/l 6.7

The enzyme concentrations are specified here in terms of the equivalentconcentration of the pure, catalytically active enzyme proteins, inorder to avoid ambiguity resulting from activity assay procedures.Standard biochemical techniques can be used to purify and characterisethe enzymes.

A suitable pectate lyase can be obtained from Bacillus licheniformis, asdescribed in U.S. Pat. No. 6,124,127.

The endo-glucanase is an endo-glucanase with anti-redeposition effect asdefined above. A method for preparation of this enzyme is described inExample 2.

In this test the soil contains a pectin. The results show that thecombinations of the endo-glucanase and pectate lyase give a higherenzyme detergency benefit than the pectate lyase alone.

EXAMPLE 11 Wash test: Lipex® and Endo-Glucanase

The purpose of this test was to measure the enzyme detergency benefit ofa lipase (Lipex®) alone and in combination with endo-glucanase on a soilthat contains triglyceride oils.

Lipex® is the trade name of a commercially available lipase productproduced by Novozymes A/S. Lipex® is intended for use in detergents forlaundry and hard surface cleaning.

For this test the soiled swatches were prepared as follows: Swatches ofthe prewashed #2003 fabric, prepared as above, were soiled as follows. Asuspension of 1.5 g of oat flakes (rolled, toasted oats, containingapproximately 7% fats by weight, e.g. from Faellesindkob I/S, 2605Brondby, Denmark) was prepared in 500 ml tap water by stirring andheating to boiling and homogenising with an UltraTurrax® blender andthen cooling. To 100 ml of this suspension 166 mg of carbon black(“carbon for detergency tests”, supplied by Sentakukagaku-kyokai, 2-11-1Shimomaruko Ohta-ku, Tokyo 146-8620, Japan) were added, and thesuspension was homogenised with an UltraTurrax blender. 25 ml of cornoil (refined corn oil, for household use, 100% oil by weight) were addedand the mixture was homogenised with an UltraTurrax® blender. The cottonswatches were placed flat on a horizontal metal surface. 100 μl of thecarbon/oil suspension was pipetted onto the centre of each cottonswatch. The soiled cotton swatches were allowed to dry at roomtemperature overnight.

For this test the detergent composition was as described above and withthe following surfactants and pH:

-   -   LAS: 0.4 g per liter    -   Nonionic: 0.29 per liter    -   pH: 9.5

Enzyme detergency benefit results: Lipex ® at 140 μg/l Lipex ® at 710μg/l No endo-glucanase −1.7 −4.0 Endo-glucanase at 75 μg/l 46.4Endo-glucanase at 190 μg/l 53.9

The enzyme concentrations are specified here in terms of the equivalentconcentration of the pure, catalytically active enzyme proteins, inorder to avoid ambiguity resulting from activity assay procedures.Standard biochemical techniques can be used to purify and characterisethe enzymes.

The lipase can either be obtained from samples of the commercial productLipex® from Novozymes A/S or derived from suitable Humicola lanuginosavariants as described in WO 00/60063.

The endo-glucanase is an endo-glucanase with anti-redeposition effect asdefined above. A method for preparation of this enzyme is described inExample 2.

In this test the soil contains a triglyceride oil. The results show thatthe combinations of the endo-glucanase and Lipex® give a higher enzymedetergency benefit than Lipex® alone.

EXAMPLE 12 Tests for Endo-Glucanase with Anti-Redeposition Effect

Two enzymes were evaluated by the tests for “endo-glucanase withanti-redeposition effect”. These were:

-   -   The enzyme that can be prepared as described in Example 2 above        (“Example 2 enzyme”), and    -   Carezyme, i.e. the 43 kD cellulase derived from Humicola        insolens.

Results: deltaOD from test for endo-glucanase activity Anti-redepositioneffect Example 2 enzyme 0.4 10 Carezyme >1 0

The results show that the Example 2 enzyme is an anti-redepositionendo-glucanase, i.e. an endo-glucanase with anti-redeposition effectaccording to the invention, and that Carezyme is not.

1. A detergent composition comprising an endo-glucanase, wherein theendo-glucanase is selected from one of: (i) the endo-glucanase havingthe amino acid sequence of position 1 to position 773 of SEQ ID NO: 2;(ii) an endo-glucanase having a sequence of at least 90% identity to theamino acid sequence of position 1 to position 773 of SEQ ID NO:2; or afragment thereof that has glucanase activity, when identity isdetermined by GAP provided in the GCG program package using a GAPcreation penalty of 3.0 and GAP extension penalty of 0.1.
 2. A detergentcomposition comprising an endo-glucanase, wherein the endo-glucanase isan anti-redeposition endo-glucanase as determined by the test forendo-glucanase activity together with the test for anti-redepositioneffect.
 3. A detergent composition comprising anionic tensides and acombination of an endo-glucanase as described in claims 1 and a fungalcellulase, wherein both enzymes are stable in the presence of anionictensides.
 4. The detergent composition of claim 3, wherein (a) theendo-glucanase is selected from one of: (i) the endo-glucanase havingthe amino acid sequence of position 1 to position 773 of SEQ ID NO: 2;(ii) an endo-glucanase having a sequence of at least 90% identity to theamino acid sequence of position 1 to position 773 of SEQ ID NO:2; or afragment thereof that has glucanase activity, when identity isdetermined by GAP provided in the GCG program package using a GAPcreation penalty of 3.0 and GAP extension penalty of 0.1; (b) thecellulase is selected from one of: (i) the cellulase having the aminoacid sequence of position 1 to position 299 of SEQ ID NO: 4 or (ii) acellulase having a sequence of at least 70% identity to the amino acidsequence of position 1 to position 299 of SEQ ID NO:4, or a fragmentthereof that has cellulase activity, when identity is determined by GAPprovided in the GCG program package using a GAP creation penalty of 3.0and GAP extension penalty of 0.1.
 5. The detergent composition of claims1, wherein the endo-glucanase is active at a pH at least in the range of4-11, preferably 5.5-10.5.
 6. The detergent composition of claims 3,wherein cellulase is derived from a strain of the genus Thielavia,preferably a strain of Thielavia terrestris, especially Thielaviaterrestris NRRL 8126 and shown in SEQ ID NO:
 4. 7-21. (canceled)
 22. Aprocess for washing a fabric, comprising contacting a fabric with anaqueous solution of a composition of claims 1, optionally underagitation, for an effective period of time.
 23. The process of claim 22,wherein the period of time is between 2 minutes and 24 hours, preferably10 minutes to 60 minutes.
 24. A process of claim 23, wherein the weightratio of the endo-glucanase protein component to the total enzymeprotein is less than 1:2.
 25. A process for washing a hard surface,comprising contacting the surface with an aqueous solution of acomposition of claims 1, for an effective period of time.
 26. Theprocess of claim 22, wherein the period of time is between 1 minute and1 hour, preferably 5 minutes to 30 minutes.
 27. A process of claims 22,wherein the weight ratio of the endo-glucanase protein component to thetotal enzyme protein is less than 1:2.